Protease inhibitors

ABSTRACT

The present invention provides compounds which inhibit proteases, including cathepsin K, pharmaceutical compositions of such compounds, and methods for treating diseases of excessive bone loss or cartilage or matrix degradation, including osteoporosis; gingival disease including gingivitis and periodontitis; arthritis, more specifically, osteoarthritis and rheumatoid arthritis; Paget&#39;s disease; hypercalcemia of malignancy; and metabolic bone disease, comprising inhibiting said bone loss or excessive cartilage or matrix degradation by administering to a patient in need thereof a compound of the present invention.

This application is a continuation of application Ser. No. 09/633,700filed Aug. 7, 2000 now abandoned; which is a continuation application ofapplication Ser. No. 09/330,284, filed Jun. 11, 1999 now abandoned;which is a divisional application of Ser. No. 08/793,915, filed on Apr.30, 1998 now abandoned; which is a 371 of International Application No.PCT/US96/18000 filed Oct. 30, 1996, which claims the benefit of priorityof Provisional Application No. 60/008,108, filed Oct. 30, 1995, No.60/007,473, filed Nov. 22, 1995, No. 60/008,992, filed Dec. 21, 1995,No. 60/013,748, filed Mar. 20, 1996, No. 60/013,764, filed Mar. 20,1996, No. 60/013,747, filed Mar. 20, 1996, No. 60/017,455, filed May 17,1996, No. 60/017,892, filed May 17, 1996, No. 60/022,047, filed Jul. 22,1996 and 60/023,494, filed Aug. 7, 1996.

FIELD OF THE INVENTION

This invention relates in general to hydrazidyl, bis-hydrazidyl andbis-aminomethyl carbonyl protease inhibitors, particularly suchinhibitors of cysteine and serine proteases, more particularly compoundswhich inhibit cysteine proteases, even more particularly compounds whichinhibit cysteine proteases of the papain superfamily, yet moreparticularly compounds which inhibit cysteine proteases of the cathepsinfamily, most particularly compounds which inhibit cathepsin K. Suchcompounds are particularly useful for treating diseases in whichcysteine proteases are implicated, especially diseases of excessive boneor cartilage loss, e.g., osteoporosis, periodontitis, and arthritis.

BACKGROUND OF THE INVENTION

Cathepsins are a family of enzymes which are part of the papainsuperfamily of cysteine proteases. Cathepsins B, H, L, N and S have beendescribed in the literature. Recently, cathepsin K polypeptide and thecDNA encoding such polypeptide were disclosed in U.S. Pat. No. 5,501,969(called cathepsin O therein). Cathepsin K has been recently expressed,purified, and characterized. Bossard, M. J., et al., (1996) J. Biol.Chem. 271, 12517-12524; Drake, F. H., et al., (1996) J. Biol. Chem. 271,12511-12516; Bromme, D., et al., (1996) J. Biol. Chem. 271, 2126-2132.

Cathepsin K has been variously denoted as cathepsin O or cathepsin O2 inthe literature. The designation cathepsin K is considered to be the moreappropriate one.

Cathepsins function in the normal physiological process of proteindegradation in animals, including humans, e.g., in the degradation ofconnective tissue. However, elevated levels of these enzymes in the bodycan result in pathological conditions leading to disease. Thus,cathepsins have been implicated as causative agents in various diseasestates, including but not limited to, infections by pneumocystiscarinii, trypsanoma cruzi, trypsanoma brucei brucei, and Crithidiafusiculata; as well as in schistosomiasis, malaria, tumor metastasis,metachromatic leukodystrophy, muscular dystrophy, amytrophy, and thelike. See International Publication Number WO 94/04172, published onMar. 3, 1994, and references cited therein. See also European PatentApplication EP 0 603 873 A1, and references cited therein. Two bacterialcysteine proteases from P. gingivallis, called gingipains, have beenimplicated in the pathogenesis of gingivitis. Potempa, J., et al. (1994)Perspectives in Drug Discovery and Design, 2, 445-458.

Cathepsin K is believed to play a causative role in diseases ofexcessive bone or cartilage loss. Bone is composed of a protein matrixin which spindle- or plate-shaped crystals of hydroxyapatite areincorporated. Type I collagen represents the major structural protein ofbone comprising approximately 90% of the protein matrix. The remaining10% of matrix is composed of a number of non-collagenous proteins,including osteocalcin, proteoglycans, osteopontin, osteonectin,thrombospondin, fibronectin, and bone sialoprotein. Skeletal boneundergoes remodelling at discrete foci throughout life. These foci, orremodelling units, undergo a cycle consisting of a bone resorption phasefollowed by a phase of bone replacement.

Bone resorption is carried out by osteoclasts, which are multinuclearcells of hematopoietic lineage. The osteoclasts adhere to the bonesurface and form a tight sealing zone, followed by extensive membraneruffling on their apical (i.e., resorbing) surface. This creates anenclosed extracellular compartment on the bone surface that is acidifiedby proton pumps in the ruffled membrane, and into which the osteoclastsecretes proteolytic enzymes. The low pH of the compartment dissolveshydroxyapatite crystals at the bone surface, while the proteolyticenzymes digest the protein matrix. In this way, a resorption lacuna, orpit, is formed. At the end of this phase of the cycle, osteoblasts laydown a new protein matrix that is subsequently mineralized. In severaldisease states, such as osteoporosis and Paget's disease, the normalbalance between bone resorption and formation is disrupted, and there isa net loss of bone at each cycle. Ultimately, this leads to weakening ofthe bone and may result in increased fracture risk with minimal trauma.

Several published studies have demonstrated that inhibitors of cysteineproteases are effective at inhibiting osteoclast-mediated boneresorption, and indicate an essential role for a cysteine proteases inbone resorption. For example, Delaisse, et al., Biochem. J., 1980, 192,365, disclose a series of protease inhibitors in a mouse bone organculture system and suggest that inhibitors of cysteine proteases (e.g.,leupeptin, Z-Phe-Ala-CHN₂) prevent bone resorption, while serineprotease inhibitors were ineffective. Delaisse, et al., Biochem.Biophys. Res. Commun., 1984, 125, 441, disclose that E-64 and leupeptinare also effective at preventing bone resorption in vivo, as measured byacute changes in serum calcium in rats on calcium deficient diets.Lerner, et al., J. Bone Min. Res., 1992, 7, 433, disclose that cystatin,an endogenous cysteine protease inhibitor, inhibits PFH stimulated boneresorption in mouse calvariae. Other studies, such as by Delaisse, etal., Bone, 1987, 8, 305, Hill, et al., J. Cell. Biochem., 1994, 56, 118,and Everts, et al., J. Cell. Physiol., 1992, 150, 221, also report acorrelation between inhibition of cysteine protease activity and boneresorption. Tezuka, et al., J. Biol. Chem., 1994, 269, 1106, Inaoka, etal., Biochem. Biophys. Res. Commun., 1995, 206, 89 and Shi, et al., FEBSLett., 1995, 357, 129 disclose that under normal conditions cathepsin K,a cysteine protease, is abundantly expressed in osteoclasts and may bethe major cysteine protease present in these cells.

The abundant selective expression of cathepsin K in osteoclasts stronglysuggests that this enzyme is essential for bone resorption. Thus,selective inhibition of cathepsin K may provide an effective treatmentfor diseases of excessive bone loss, including, but not limited to,osteoporosis, gingival diseases such as gingivitis and periodontitis,Paget's disease, hypercalcemia of malignancy, and metabolic bonedisease. Cathepsin K levels have also been demonstrated to be elevatedin chondroclasts of osteoarthritic synovium. Thus, selective inhibitionof cathepsin K may also be useful for treating diseases of excessivecartilage or matrix degradation, including, but not limited to,osteoarthritis and rheumatoid arthritis. Metastatic neoplastic cellsalso typically express high levels of proteolytic enzymes that degradethe surrounding matrix. Thus, selective inhibition of cathepsin K mayalso be useful for treating certain neoplastic diseases.

Several cysteine protease inhibitors are known. Palmer, (1995) J. Med.Chem., 38, 3193, disclose certain vinyl sulfones which irreversiblyinhibit cysteine proteases, such as the cathepsins B, L, S, O2 andcruzain. Other classes of compounds, such as aldehydes, nitrites,α-ketocarbonyl compounds, halomethyl ketones, diazomethyl ketones,(acyloxy)methyl ketones, ketomethylsulfonium salts and epoxy succinylcompounds have also been reported to inhibit cysteine proteases. SeePalmer, id, and references cited therein.

U.S. Pat. No. 4,518,528 discloses peptidyl fluoromethyl ketones asirreversible inhibitors of cysteine protease. Published InternationalPatent Application No. WO 94/04172, and European Patent Application Nos.EP 0 525 420 A1, EP 0 603 873 A1, and EP 0 611 756 A2 describealkoxymethyl and mercaptomethyl ketones which inhibit the cysteineproteases cathepsins B, H and L. International Patent Application No.PCT/US94/08868 and and European Patent Application No. EP 0 623 592 A1describe alkoxymethyl and mercaptomethyl ketones which inhibit thecysteine protease IL-1β convertase. Alkoxymethyl and mercaptomethylketones have also been described as inhibitors of the serine proteasekininogenase (International Patent Application No. PCT/GB91/01479).

Azapeptides which are designed to deliver the azaamino acid to theactive site of serine proteases, and which possess a good leaving group,are disclosed by Elmore et al., Biochem. J., 1968, 107, 103, Garker etal., Biochem. J., 1974, 139, 555, Gray et al., Tetrahedron, 1977, 33,837, Gupton et al., J. Biol. Chem., 1984, 259, 4279, Powers et al., J.Biol. Chem., 1984, 259, 4288, and are known to inhibit serine proteases.In addition, J. Med. Chem., 1992, 35, 4279, discloses certain azapeptideesters as cysteine protease inhibitors.

Antipain and leupeptin are described as reversible inhibitors ofcysteine protease in McConnell et al., J. Med. Chem., 33, 86; and alsohave been disclosed as inhibitors of serine protease in Umezawa et al.,45 Meth. Enzymol. 678. E64 and its synthetic analogs are also well-knowncysteine protease inhibitors (Barrett, Biochem. J., 201, 189, andGrinde, Biochem. Biophys. Acta, 701, 328). U.S. Pat. No. 5,142,056describes 1,3-diamido-propanones which inhibit HIV protease.1,3-diamido-propanones have also been described as analgesic agents(U.S. Pat. Nos. 4,749,792 and 4,638,010).

Certain heterocyclic derivatives of amino acids have been disclosed inthe art. For instance, Hamada, et al., PEPTIDE CHEMISTRY, 1983.Proceedings of the 21st Symposium on Peptide Chemistry (1984), andBoden, et al., Tet. Lett., 1994, 35, 8271 (1994) disclose thiazolederivatives; and Borg, et al., 1995, 60, 3112, disclose oxadiazole andtriazole derivatives.

The synthesis of azatides (polyacylhydrazides) as peptide mimetics hasrecently been disclosed by Han and Janda, J. Am. Chem. Soc. 1996, 118,2539.

Thus, a structurally diverse variety of cysteine protease inhibitorshave been identified. However, these known inhibitors are not consideredsuitable for use as therapeutic agents in animals, especially humans,because they suffer from various shortcomings. These shortcomingsinclude lack of selectivity, cytotoxicity, poor solubility, and overlyrapid plasma clearance. A need therefore exists for methods of treatingdiseases caused by pathological levels of cysteine proteases, includingcathepsins, especially cathepsin K, and for novel inhibitor compoundsuseful in such methods.

We have now discovered a novel class of hydrazidyl, bis-hydrazidyl andbis-aminomethyl carbonyl compounds which are protease inhibitors, mostparticularly of cathepsin K.

SUMMARY OF THE INVENTION

An object of the present invention is to provide hydrazidyl,bis-hydrazidyl and bis-aminomethyl carbonyl protease inhibitors,particularly such inhibitors of cysteine and serine proteases, moreparticularly such compounds which inhibit cysteine proteases, even moreparticularly such compounds which inhibit cysteine proteases of thepapain superfamily, yet more particularly such compounds which inhibitcysteine proteases of the cathepsin family, most particularly suchcompounds which inhibit cathepsin K, and which are useful for treatingdiseases which may be therapeutically modified by altering the activityof such proteases.

Accordingly, in the first aspect, this invention provides a compoundaccording to Formula I.

In another aspect, this invention provides a pharmaceutical compositioncomprising a compound according to Formula I and a pharmaceuticallyacceptable carrier, diluent or excipient.

In yet another aspect, this invention provides a method of treatingdiseases in which the disease pathology may be therapeutically modifiedby inhibiting proteases, particularly cysteine and serine proteases,more particularly cysteine proteases, even more particularly cysteineproteases of the papain superfamily, yet more particularly cysteineproteases of the cathepsin family, most particularly cathepsin K.

In a particular aspect, the compounds of this invention are especiallyuseful for treating diseases characterized by bone loss, such asosteoporosis and gingival diseases, such as gingivitis andperiodontitis, or by excessive cartilage or matrix degradation, such asosteoarthritis and rheumatoid arthritis.

DETAILED DESCRIPTION

The present invention provides compounds of Formula I:

L=C₂₋₆alkyl, Ar—C₀₋₆alkyl, Het-C₀₋₆alkyl, CH(R⁶⁶)NR⁶⁰R⁶⁸, CH(R⁶⁶)Ar,CH(R⁶⁶)OAr′, NR⁶⁶R⁶⁷;

M=C(O), SO₂;

G=

J=C(O), SO₂;

T=Ar, Het;

V=C₃₋₇cycloalkyl;

W═H, —CN, —CF₃, —NO₂, —COR⁷, —CO₂R⁶, —CONHR⁶, —SO₂NHR⁶, —NHSO₂R⁶,—NHCOR⁷, —O—COR⁶, —SR⁶, NR′R⁶, NR′(C═NH)NHR⁵, Cl, Br, I, F;

X═Y═Z═N, O, S or CR⁴,

 provided that at least two of X, Y and Z are heteroatoms and at leastone of X, Y and Z is N, or one of X, Y and Z is C═N, C═C or N═N and theother two are CR⁴ or N, provided that X, Y and Z together comprise atleast two N;

indicates a single or double bond in the five-membered heterocycle;

m=0, 1, 2;

n=1 to 6;

φ=0, 1, 2;

Ar=phenyl, naphthyl, optionally substituted by one or more ofPh-C₀₋₆alkyl, Het-C₀₋₆alkyl, C₁₋₆alkoxy, Ph-C₀₋₆alkoxy, Het-C₀₋₆alkoxy,OH, (CH₂)₁₋₆NR⁵⁸R⁵⁹, O(CH₂)₁₋₆NR⁵⁸R⁵⁹;

Ar′=phenyl or naphthyl, optionally substituted by one or more ofPh-C₀₋₆alkyl, Het-C₀₋₆alkyl, C₁₋₆alkoxy, Ph-C₀₋₆alkoxy, Het-C₀₋₆alkoxy,OH, (CH₂)₁₋₆NR⁵⁸R⁵⁹, O(CH₂)₁₋₆NR⁵⁸R⁵⁹, or halogen;

R′=H, C₁₋₆alkyl, Ar—C₀₋₆alkyl, Het-C₀₋₆alkyl;

R¹=H, C₁₋₆alkyl;

R²=C₄₋₆alkyl, C₄₋₆alkenyl, benzyl;

R³=C₁₋₆alkyl, Ar—C₀₋₆alkyl, Het-C₀₋₆alkyl, R⁵CO—, R⁵SO₂—, R⁵OC(O)—,R⁵NHCO—;

R⁴=H, C₁₋₆alkyl, Ar—C₀₋₆alkyl, Het-C₀₋₆alkyl;

R⁵=Ar-₀₋₆alkyl, Het-C₀₋₆alkyl;

R⁶=H, C₁₋₆alkyl, CH₂CF₃, Ar—C₀₋₆alkyl, Het-C₀₋₆alkyl;

R⁷=C₁₋₆alkyl, Ar—C₀₋₆alkyl, Het-C₀₋₆alkyl;

R⁸=H; C₂₋₆ alkenyl; C₂₋₆alkynyl; Het; Ar; C₁₋₆alkyl, optionallysubstituted by OR′, SR′, NR′₂, CO₂R′, CO₂NR′₂, N(C═NH)NH₂, Het or Ar;

R⁹=H, C₁₋₆alkyl, Ar—C₀₋₆alkyl, Het-C₀₋₆alkyl;

R¹⁰=C₁₋₆alkyl, Ar—C₀₋₆alkyl, Het-C₀₋₆alkyl;

R¹¹=H, C₁₋₆alkyl, Ar—C₁₋₆alkyl, Het-C₀₋₆alkyl, or

R¹²=H, C₁₋₆alkyl, Ar—C₀₋₆alkyl, Het-C₀₋₆alkyl;

R¹³=H, C₁₋₆alkyl, Ar—C₀₋₆alkyl, Het-C₀₋₆alkyl;

R¹⁵=H, C₁₋₆alkyl, C₂₋₆alkeny), C₂₋₆alkynyl, Ar, Het, or C₁₋₆alkyloptionally substituted by OR⁹, NR⁹ ₂, CONR⁹ ₂, N(C═NH)NH—, Het or Ar;

R¹⁶=C₂₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl, Ar, Het, or C₂₋₆alkyloptionally substituted by OR⁹, SR⁹, NR⁹ ₂, CO₂R⁹, CONR⁹ ₂, N(C═NH)NH—,Het or Ar;

R¹⁹=H, C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl, Ar, Het, or C₁₋₆alkyloptionally substituted by OR⁹, SR⁹, NR⁹ ₂, CO₂R⁹, CONR⁹ ₂, N(C═NH)NH—,Het or Ar;

R¹⁷=R⁷²=H, C₁₋₆alkyl, R¹⁰, R¹⁰C(O)—, R¹⁰C(S)—, R¹⁰OC(O)—;

R²¹=R²⁶=C₅₋₆alkyl; C₂₋₆alkenyl; C₃₋₁₁cycloalkyl; T—C₃₋₆alkyl;V—C₁₋₆alkyl; T—C₂₋₆alkenyl; T—(CH₂)_(n)CH(T)(CH₂)_(n); optionallysubstituted by one or two halogens, SR²⁰, OR²⁰,NR²OR²⁷ or C₁₋₄alkyl;

R²⁷=R²⁸CO, R²⁸OCO;

R²⁸=C₁₋₆alkyl; C₃₋₁₁cycloalkyl; Ar; Het; T—C₁₋₆alkyl;T—(CH₂)_(n)CH(T)(CH₂)_(n); optionally substituted by one or twohalogens, SR²⁰, OR²⁰, NR²OR⁷³, C₁₋₆alkyl;

R²⁰=R²²=R²³=R²⁴=R²⁵=R⁷³=H, C₁₋₄alkyl, Ar—C₀₋₆alkyl, Het-C₀₋₆alkyl;

 Cbz-leucinyl-; 2-, 3-, or 4-pyridyl methyloxycarbonyl-leucinyl-;4-imidazole aceryl-leucinyl-, phenyl acetyl-leucinyl,N,N-dimethyl-glycinyl leucinyl, 4-pyridyl acetyl-leucinyl, 2-pyridylsulfonyl-leucinyl, 4-pyridyl carbonyl-leucinyl, acetyl-leucinyl,benzoyl-leucinyl, 4-phenoxybenzoyl-, 2- or 3-benzyloxybenzoyl-, biphenylacetyl, alpha-isobutyl-biphenyl acetyl, Cbz-phenylaaninyl,Cbz-norleucinyl-, Cbz-norvalinyl-, Cbz-glutamyl-, Cbz-epsilon-(t-butylester)-glutarnyl; acetyl-leucinyl-, 6- or 8-quinoline carbonyl, biphenylacetyl, alpha-isobutyl-biphenyl acetyl, acetyl, benzoyl, 2- or3-benzyloxy benzoyl, 4-phenoxy benzoyl-,

 Cbz-amino acid-; 2-,3-, or 4-pynidylmethyloxyearbonyl-aminoacid-; arylC₀-C₆alkyloxy carbonyl-amino acid- , heteroaryl C₀-C₆alkyloxycarbonyl-amino acid-, aryl C₀-C₆alkyloxy carbonyl-amino acid-,heteroaryl C₀-C₆alkyloxy carbonyl-amino acid-, C₁-C₆alkyloxycarbonyl-amino acid-; C₁-C₆alkyl carbonyl, aryl C₀-C₆alkyl carbonyl,heteroaryl C₀-C₆alkyl carbonyl, aryl C₀-C₆alkyl carbonyl, heteroarylC₀-C₆alkyl carbonyl, C₁-C₆alkyl sulfonyl, aryl C₀-C₆alkyl sulfonyl,heteroaryl C₀-C₆alkyl sulfonyl, aryl C₀-C₆alkyl sulfonyl, heteroarylC₀-C₆alkyl sulfonyl;

R³⁰=—H, C₁₋₆ alkyl;

 Cbz-leucinyl-; 2-, 3-, or 4-pyridyl methyloxycarbonyl-leucinyl-;4-imidazole acetyl-leucinyl-, phenyl acetyl-leucinyl,N,N-dimethyl-glycinyl leucinyl, 4-pyridyl acetyl-leucinyl, 2-pyridylsulfonyl-leucinyl, 4-pyridyl carbonyl-leucinyl, acetyl-leucinyl,benzoyl-leucinyl, 4-phenoxybenzoyl-, 2- or 3-benzyloxybenzoyl-, biphenylacetyl, alpha-isobutyl-biphenyl acetyl, Cbz-phenylalaninyl,Cbz-norleucinyl-, Cbz-norvalinyl-, Cbz-glutamyl-, Cbz-epsilon-(t-butylester)-glutamyl; acetyl-leucinyl-, 6- or 8-quinoline carbonyl, biphenylacetyl, alpha-isobutyl-biphenyl acetyl, acetyl, benzoyl, 2- or3-benzyloxy benzoyl, 4-phenoxy benzoyl-,

 Cbz-amino acid-; 2-,3-, or 4-pyridylmethyloxycarbonyl-aminoacid-; arylC₀-C₆alkyloxy carbonyl-amino acid-, heteroaryl C₀-C₆alkyloxycarbonyl-amino acid-, aryl C₀-C₆alkyloxy carbonyl-amino acid-,heteroaryl C₀-C₆alkyloxy carbonyl-amino acid-, C₁-C₆alkyloxycarbonyl-amino acid-; C₁-C₆alkyl carbonyl, aryl C₀-C₆alkyl carbonyl,heteroaryl C₀-C₆alkyl carbonyl, aryl C₀-C₆alkyl carbonyl, heteroarylC₀-C₆alkyl carbonyl, C₁-C₆alkyl sulfonyl, aryl C₀-C₆alkyl sulfonyl,heteroaryl C₀-C₆alkyl sulfonyl, aryl C₀-C₆alkyl sulfonyl, heteroarylC₀-C₆alkyl sulfonyl;

R³²OCH₂Ar, OCH₂C₁₋₆alkyl, aryl substituted C₀₋₆alkyl, heteroarylsubstituted C₀₋₆alkyl,4-imidazole methylene; 2-, 3-, or4-pyridylmethylneneoxy; 4-pyridyl methylene, 2-pyridyl sulfonyl,4-pyridyl, aryl substituted C₀₋₆alkyloxy, heteroaryl substitutedC₀₋₆alkyloxy;

R³³=C₁₋₆alkyl, —CH₂Ph, —CH₂CH₂CO₂R³⁴;

R³⁴=—H, C₁₋₆alkyl;

R³⁵=Ar, HetAr;

R³⁶=Aryl, heteroaryl, pyridyl, isoquinolinyl;

R³⁷=C₁₋₆alkyl, —CH₂Ph, —CH₂CH₂CO₂R³⁴;

R³⁸=Cbz; C₁₋₆alkyl or aryl substituted

 Cbz; C₁₋₆alkyl —CO; benzoyl; C₁₋₆alkyl or aryl substituted benzoyl;

 Cbz-leucinyl-; 2-, 3-, or 4-pyridyl methyloxycarbonyl-leucinyl-;4imidazole acetyl-leucinyl-, phenyl acetyl-leucinyl,N,N-dimethyl-glycinyl leucinyl, 4-pyridyl acetyl-leucinyl, 2-pyridylsulfonyl-leucinyl, 4pyridyl carbonyl-leucinyl, acetyl-leucinyl,benzoyl-leucinyl, 4-phenoxybenzoyl-, 2- or 3-benzyloxybenzoyl-, biphenylacetyl, alpha-isobutyl-biphenyl acetyl, Cbz-phenylalaninyl,Cbz-norleucinyl-, Cbz-norvalinyl-, Cbz-glutarnyl-, Cbz-epsilon-(t-butylester)-glutamyl; acetyl-leucinyl-, 6- or 8-quinoline carbonyl, biphenylacetyl, alpha- isobutyl-biphenyl acetyl, acetyl, benzoyl, 2- or3-benzyloxy benzoyl, 4-phenoxy benzoyl-,

 Cbz-amino acid-; 2-,3-, or 4-pyridylmethyloxycarbonyl-aminoacid-; arylC₀-C₆alkyloxy carbonyl-amino acid-, heteroaryl C₀-C₆alkyloxycarbonyl-amino acid-, aryl C₀-C₆alkyloxy carbonyl-amino acid-,heteroarylC₀-C₆alkyloxy carbonyl-amino acid-, C₁-C₆alkyloxy carbonyl-amino acid-;C₁-C₆alkyl carbonyl, aryl C₀-C₆alkyl carbonyl, heteroaryl C₀-C₆alkylcarbonyl, aryl C₀-C₆alkyl carbonyl, heteroaryl C₀-C₆alkyl carbonyl,C₁-C₆alkyl sulfonyl, aryl C₀-C₆alkyl sulfonyl, heteroaryl C₀-C₆alkylsulfonyl, aryl C₀-C₆alkyl sulfonyl, heteroaryl C₀-C₆alkyl sulfonyl;

R⁴⁰=H and C₁₋₆alkyl;

R⁴¹=H and C₁₋₆alkyl;

R⁴²=C₁₋₆alkyl, aryl substituted C₁₋₆alkyl and hetero aryl substitutedC₁₋₆alkyl,; H when R⁴³ is C₁₋₆alkyl, aryl substituted C₁₋₆alkyl; andheteroaryl substituted C₁₋₆alkyl;

R⁴³=C₁₋₆alkyl, aryl substituted C₁₋₆alkyl and hetero aryl substitutedC₁₋₆alkyl,; H when R⁴² is C₁₋₆alkyl, aryl substituted C₁₋₆alkyl; andheteroaryl substituted C₁₋₆alkyl;

R⁴⁴=CH(R⁵³)NR⁴⁵R⁵⁴, CH(R⁵⁵)Ar, C₅₋₆alkyl;

R⁴⁵=R⁴⁶=R⁴⁷=R⁴⁸=R⁴⁹=R⁵⁰=R⁵¹=H, C₁₋₆alkyl, Ar—C₀₋₆alkyl, Het-C₀₋₆alkyl;

R⁵²=Ar, Het, CH(R⁵⁶)Ar, CH(R⁵⁶)OAr, N(R⁵⁶)Ar, C₁₋₆alkyl, CH(R⁵⁶)NR⁴⁶R⁵⁷;

R⁵³=C₂₋₆alkyl, Ar—C₀₋₆alkyl, Het-C₀₋₆alkyl,

 R⁵³ and R⁴⁵ may be connected to form a pyrrolidine or piperidine ring;

R⁵⁴=R⁵⁷=R⁴⁷, R⁴⁷C(O), R⁴⁷C(S), R⁴⁷OC(O);

R⁵⁵=R⁵⁶=R⁵⁸=R⁵⁹=H, C₁₋₆alkyl, Ar—C₀₋₆alkyl, Het-C₀₋₆alkyl;

R⁶⁰=R⁶¹=R⁶²=R⁶³=R⁶⁴=H, C₁₋₆alkyl, Ar—C₀₋₆alkyl, or Het-C₀₋₆alkyl;

R⁶⁵=C₁₋₆alkyl, Ar, Het, CH(R⁶⁹)Ar, CH(R⁶⁹)OAr, N(R⁶⁹)Ar, CH(R⁶⁹)NR⁶¹R⁷⁰;

R⁶⁶=R⁶⁹=R⁷¹=H, C₁₋₆alkyl, (CH₂)₀₋₆—C₃₋₆cycloalkyl, Ar—C₀₋₆alkyl,Het-C₀₋₆alkyl;

R⁶⁷=C₁₋₆alkyl, (CH₂)₀₋₆—C₃₋₆cycloalkyl, Ar—C₀₋₆alkyl, Het-C₀₋₆alkyl; R⁶⁶and R⁶⁷ may be combined to form a 3-7 membered monocyclic or7-10-membered bicyclic carbocyclic or heterocyclic ring, optionallysubstituted with 1-4 Of C₁₋₆alkyl, Ph-C₀₋₆alkyl, Het-C₀₋₆alkyl,C₁₋₆alkoxy, Ph-C₀₋₆alkoxy, Het-C₀₋₆alkoxy, OH, (CH₂)₁₋₆NR⁵⁸R⁵⁹,O(CH₂)₁₋₆NR⁵⁸R⁵⁹;

R⁶⁸=R⁷⁰=R⁶², R⁶²C(O), R⁶²C(S), R⁶²OC(O), R⁶²OC(O)NR⁵⁹CH(R⁷¹)(CO);

and pharmaceutically acceptable salts thereof.

The compounds of Formula I are hydrazidyl, bis-hydrazidyl andbis-aminomethyl carbonyl compounds having in common key structuralfeatures required of protease substrates, most particularly cathepsin Ksubstrates. These structural features endow the present compounds withthe appropriate molecular shape necessary to fit into the enzymaticactive site, to bind to such active site, and to react with a sulfhydrylgroup on the active site, thereby blocking the site and inhibitingenzymatic biological activity. Referring to Formula I, such structuralfeatures include the central electrophilic carbonyl, a peptidyl orpeptidomimetic molecular backbone on either side of the centralcarbonyl, a terminal carbobenzyloxy moiety (e.g., Cbz-leucinyl), or amimic thereof, on the backbone on one or both sides of the carbonyl, andoptionally, an isobutyl side chain extending from the backbone on one orboth sides of the carbonyl.

Compounds of Formula I wherein D=

and Q=

are preferred embodiments of the present invention. For the sake ofconvenience, such compounds are referred to herein after as compounds ofFormula II.

More preferred embodiments of the present invention include compounds ofFormula II wherein:

X=S, Y=CH, and Z=N;

X=CH, Y=S, and Z=N;

X=N, Y=N, and Z=S;

X=N, Y=N, and Z=O; and

X=N, Y=N, and Z=N.

Preferably R¹ is H, methyl or isobutyl. Preferably R¹ is isobutyl.

Preferably R² is isobutyl or benzyl.

Preferably R³ is R⁵OC(O)—, particularly benzyloxycarbonyl.

Preferably A is a D- or L-amino acid or is absent, preferably A isabsent.

Preferably W is CN, NHR⁶, SR⁶, CONHR⁶ or CO₂R⁶. Suitably R⁶ is H,C₁₋₄alkyl, phenyl or benzyl. Typically, W is CO₂H, CO₂—C₁₋₄alkyl,CO₂-Ph, CO₂—CH₂Ph, CONH₂, NH₂ or SH.

The following components or formula II are particularly preferred:

(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-carboxythiazol-4-yl)-3′-methylbutyl]-4-methylpentanamide;

(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-carboxamidothiazol-4-yl)-3′-methylbutyl]4-methylpentanamide;

(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-carboethoxythiazol-4-yl)-3′-methylbutyl]4-methylpentanamide;

(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-cyanothiazol-4-yl)-3′-methylbutyl]-4-methylpentanamide;

(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-[2-(N′-benzylcarboxamido)thiazol-4-yl]-3′-methylbutyl]4-methylpentanamide;

(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-[2-[N′-(3-methylpropyl)carboxamido]thiazol-4-yl)]-3′-methylbutyl]4-methylpentanamide;

(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-[2-[N′-(2-phenylethyl)carboxamido]thiazol-4yl)]-3′-methylbutyl]4-methylpentanamide;

(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(4-carboethoxythiazol-2-yl)-3′-methylbutyl]-4-methylpentanamide;

(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(4-carboxythiazol-2-yl)-3′-methylbutyl]4-methylpentanamide;

(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(4carboethoxythiadiazol-2-yl)-3′-methylbutyl]4-methylpentanamide;

(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-carbo-2,2,2-trifluoroethoxythiazol-4-yl)-3′-methylbutyl]4-methylpentanamide;

(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(4-carboetboxyoxadiazol-2-yl)-3′-methylbutyl]4-methylpentanamide;

(2S,1′S)-2-(benzyloxycarbonyl-L-leucinyl)amino-N-[1′-(4-carboethoxythiazol-2-yl)-3′-methylbutyl]4-methylpentanamide;

(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(4-carboxamidooxadiazol-2-yl)-3′-methylbutyl]-4-methylpentanamide;

(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-carboethoxythiazol-4-yl)-3′-methylbutyl]-3-phenylpropanamide;

(2S,1′S)-2-(benzyloxycarbonyl-L-leucinyl)amino-N-[1′-(2-carboethoxythiazol-4-yl)-3′-methylbutyl]-4-methylpentanamide;

(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(5-mercapto-1,2,4-oxadiazol-3-yl)-3′-methylbutyl]4-methylpentanamide;

(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-mercaptothiazol-4-yl)-3′-methylbutyl]4-methylpentanamide;

(2S)-2-(benzyloxycarbonyl)amino-N-(4-carboethoxythiazol-2-yl)methyl-4-methylpentanamide;

(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-benzyloxycarbonylthiazol-4-yl)-3′methylbutyl]-4-methylpentanamide;

(2S,1′S)-2-(benzyloxycarbonyl)amino-4-methyl-N-[3′-methyl-1′-(2-phenoxycarbonylthiazol-4-yl)butyl]pentanamide;

(2S,1′S)-2-(benzyloxycarbonyl)amino-4-methyl-N-[3′-methyl-1′-[2-(2-methylpropyloxycarbonyl)thiazol-4-yl]butyl]pentanamide;

(2R,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(4-carboethoxythiazol-2-yl)ethyl]-4-methylpentanamide;

(2R,1′R)-2-(benzyloxycarbonyl)amino-N-[1′-(4-carboethoxythiazol-2-yl)ethyl]-4-methylpentanamide;and

(2S,1′S)-N-[1′-(2-aminothiazol-4-yl)-3′-methylbutyl]-2-(benzyloxycarbonyl)amino-4-methylpentanamide.

Most particularly preferred compounds of Formula II are:

(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-carboethoxythiazol-4-yl)-3′-methylbutyl]-4-methylpentanamide;

(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(4-carboethoxythiazol-2-yl)-3′methylbutyl]4methylpentanamide;and

2S,1′S)-2-(benzyloxycarbonyl-L-leucinyl)amino-N-[1′-(4-carboethoxythiazol-2-yl)-3′-methylbutyl]-4-methylpentanamide.

Compounds of Formula I wherein D=^(R11B—) and Q=

are preferred embodiments of the present invention. For the sake ofconvenience, such compounds are referred to herein after as compounds ofFormula III.

With respect to compounds of Formula m:

Preferably B is

More preferably B is

Preferably R¹¹ is

Preferably R¹² and R¹³ are H.

Preferably R¹⁴ is

Preferably R¹⁵, R¹⁶, R¹⁸ and R¹⁹ are C₁₋₆alkyl.

More preferably R¹⁵ and R¹⁸ are C₄₋₆alkyl.

Preferably Ar is phenyl optionally substituted by one or two groupschosen from halogen, CF₃, NO₂, SR⁹, OR⁹, NR⁹ or C₁₋₄alkyl.

Preferably R¹⁷ and R⁷² are R¹⁰OC(O)—; and more preferably R¹⁰ isAr—C₁₋₄alkyl.

Preferably, R¹⁶ and R¹⁹ are C₄₋₆alkyl; more preferably, R¹⁶ and R¹⁹ arei-Bu.

Preferably R¹⁷ and R⁷² are Cbz.

One particular embodiment of the invention of Formula III is a compoundof Formula F:

wherein X, Y, Z, R¹⁶, R¹⁷, R¹⁹ and R⁷² are as described in Formula III.

Most particularly preferred compounds of Formula III are:

(1S)-N-[4[(1-benzyloxycarbonylamino)-3-methylbutyl]thiazol-2-ylcarbonyl]-N′-(N-benzyloxycarbonyl-L-leucinyl)hydrazide;

N-benzyloxycarbonyl-L-leucinyl-N′-benzyloxycarbonyl-L-leucinyl-L-leucinylhydrazide;and

(1S)-N-[2-[(1-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-(N-benzyloxycarbonyl-L-leucinyl)hydrazide.

Compounds of Formula I wherein D=

and Q=

are preferred embodiments of the present invention. For the sake ofconvenience, such compounds are referred to herein after as compounds ofFormula IV.

A more preferred embodiment of the present invention is a compound ofFormula IV wherein R²¹ and R²⁶ are selected from the group consistingof:

N-Cbz-leucinyl, N-Cbz-glycinyl, N-acetyl-leucinyl, N-Cbz-alanyl,

and R²², R²³, R²⁴ and R²⁵ are H.

Particularly preferred embodiments of Formula IV are:

2,2′-(N,N′-bis-benzyloxycarbonyl-L-leucinyl)carbohydrazide;

2,2′-(N,N′-bis-cyclohexylacetyl)carbohydrazide;

2,2′-(N,N′-bis-4-methylpentanoyl)carbohydrazide;

2,2′-(N,N′-bis-cyclopentylacetyl)carbohydrazide,

2,2′-(N,N′-bis-benzyloxycarbonylglycinyl)carbohydrazide;

2,2′-(N,N′-bis-acetyl-L-leucinyl)carbohydrazide;

2,2′-(N,N′-bis-benzyloxycarbonyl-L-alanyl)carbohydrazide; and

2-(N-benzyloxycarbonyl-L-leucinyl)-2′-[N′-(4-methylpentanoyl)]carbohydrazide.

2,2′-(N,N′-bis-benzyloxycarbonyl-L-leucinyl)carbohydrazide is a mostpreferred embodiment of Formula IV.

Compounds of Formula I wherein D=

and Q=

are preferred embodiments of the present invention. For the sake ofconvenience, such compounds are referred to herein after as compounds ofFormula V.

In more preferred compounds of Formula V, when R³⁰=C₁-C₆ alkyl, R³⁰ ispreferably Me or —CH₂CH₂Me₂. When R³³=C₁-C₆ alkyl, R³³ is preferably—Pr, —Bu, or —CH₂CH₂Me₂. When R³⁴=C₁-C₆ alkyl, R³⁴ is preferably -t-Bu.

Even more preferred embodiments of Formula V include:

bis-(Cbz-leucinyl)-1,3-diamino-propan-2-one;

bis-1,3-(4-phenoxy-benzoyl)-diamino-propan-2-one;

1-(Cbz-leucinyl)-amino-3-(acetyl-leucinyl)-amino-propan-2-one;

1-(Cbz-leucinyl)-amino-3-(Cbz-glutanyl-t-butylester)-amino-propan-2-one;

1-(Cbz-leucinyl)-amino-3-(Cbz-glutamyl)-amino-propan-2-one;

bis-1,3-(Cbz-leucinyl)-diamino-(S)-butanone-2-one;

1-(Cbz-leucinyl)-amino-3-(Cbz-phenylalanyl)-amino-propan-2-one;

1-(Cbz-leucinyl)-amino-3-(Cbz-norleucinyl)-amino-propan-2-one;

1-(Cbz-leucinyl)-amino-3-(Cbz-norvalinyl)-amino-propan-2-one;

bis-1,3-(Cbz-leucinyl)-diamino-5-methyl-(S)-hexan-2one;

1-(acetyl-leucinyl)-amino-3-(4-phenoxy-benzoyl)-amino-propan-2-one:

1-(Cbz-homo-leucinyl)-amino-(Cbz-leucinyl)-3-amino-propan-2-one;

1-(Cbz-leucinyl)-amino-3-(acetyl-leucinyl)-amino-propan-2-one is a mostparticularly preferred embodiment of the present invention of Formula V.

Compounds of Formula I wherein D=

and Q=

are preferred embodiments of the present invention. For the sake ofconvenience, such compounds are referred to herein after as compounds ofFormula VI.

More preferably, R³⁵=Ph,

or pyridine, even more preferably, R³⁵=Ph,

Ph may be optionally substituted with C₁₋₆alkyl, C₁₋₆alkoxy, halogensand cyano groups. When R³⁵=pyridine, R may be 2-pyridyl, 3-pyridyl, or4-pyridyl.

Most particularly preferred embodiments of Formula VI include:

bis-1,3-(4-(3-chloro-2-cyano-phenoxy )-phenyl sulfonamido)-propan-2-one;

bis-1,3-(4-phenoxy-phenyl sulfonamido)-propan-2-one.

Compounds of Formula I wherein D=

and Q=

are preferred embodiments of the present invention. For the sake ofconvenience, such compounds are referred to herein after as compounds ofFormula VII.

More preferably, R³⁶ is selected from the group consisting of:

and R³⁷=Me in the more preferred compounds of Formula VII.

Particularly preferred embodiments of Formula VII are:

1-(Cbz-leucinyl)-1no-3-(4-(3-chloro-2-cyano-phenoxy)-phenylsulfonamido)-propan-2-one;

1-(Cbz-feucinyl)-amino-3-(tosyl-amino)-propan-2-one;

1-(Cbz-leucinyl)-amino-3-((4-phenoxy-phenyl)-sulfonamido)-propan-2-one;

1-(Cbz-leucinyl)-amino-3-(2-dibenzofuransulfonamido)-propan-2-one;

1-(Cbz-homo-leucinyl)-amino-3-(2-dibenzofuransulfonamido)-propan-2-one;and

1-(Cbz-leucinyl)-amino-3-(2-dibenzofuransulfonamido)-(S)-butan-2-one.

1-(Cbz-leucinyl)-amino-3-((4-phenoxy-phenyl)-sulfonamido)-propan-2-one,1-(Cbz-leucinyl)-amino-3-(2-dibenzofuransulfonamido)-propan-2-one, and1-(Cbz-leucinyl)-amino-3-(2-dibenzofuransulfonamido)-(S)-butan-2-one aremost particularly preferred embodiments of Formula VII.

Compounds of Formula I wherein D=

and Q=

are preferred embodiments of the present invention. For the sake ofconvenience, such compounds are referred to herein after as compounds ofFormula VIII.

A more preferred embodiment of Formula VIII is when R⁴³ is2-dibenzofuranylsulfonyl.

Particularly preferred embodiments of Formula VIII are:

(S)-Phenylmethyl[1-[[[3-[benzyloxycarbonyl-leucinyl-amino]-2-oxopropyl]-1-(benzyl)amino]carbonyl]-3-methylbutyl]carbamate.

(S)-Phenylmethyl[1-[[[3-[(2-dibenzofuranylsulfonyl)amino]-2-oxopropyl]-3-(benzyl)amino]carbonyl]-3-methylbutyl]carbamate

(S)-Phenylmethyl[1-[[[3-[(2-dibenzofuranylsulfonyl)amino]-2-oxopropyl]-3-(4-pyridinylmethyl)amino]carbonyl]-3-methylbutyl]carbamate

1-[[[3-[(2-dibenzofuranylsulfonyl)amino-2-oxopropyl]-3-(4-pyridinylmethyl)benzamide

(S)-Phenylmethyl [1-[[[3-[(2-dibenzofuranylsulfonyl)amino]-2-oxopropyl]-1-(4-pyridinylmethyl)amino]carbonyl]-3-methylbutyl]carbamate.

(S)-Phenylmethyl[1-[[[3-[(2-dibenzofuranylsulfonyl)amino]-2-oxopropyl]-1-(4-pyridinylmethyl)amino]carbonyl]-3-methylbutyl]carbamateis a most particularly preferred embodiment of Formula VIII.

Compounds of Formula I wherein D=

and Q=

are preferred embodiments of the present invention. For the sake ofconvenience, such compounds are referred to herein after as compounds ofFormula IX.

Compounds of Formula IX wherein:

R⁴⁴=CH(R⁵³)NHR⁵⁴;

R⁴⁵, R⁴⁶, R⁴⁸, R⁴⁹, R⁵⁰ and R⁵¹ are H;

R⁴⁷ is independently CH₃, benzyl, 2-pyridinylmethoxy,4-dimethylaminobenzyl;

J=C(O);

R⁵²=Ar, CH(R¹⁰)Ar, CH(R¹⁰)OAr, N(R¹⁰)Ar, CH(R¹⁰)NR″R¹¹;

R⁵³=ethyl, i-Bu;

R⁵⁴=R⁴⁷, R⁴⁷C(O), R⁴⁷OC(O);

R⁵⁶=H, CH₃, i-Bu;

R⁵⁷=R⁴⁷, R⁴⁷OC(O);

Ar=phenyl or naphthyl, optionally substituted by one or more ofPh-C₀₋₆alkyl, Het-C₀₋₆alkyl, C₁₋₆alkoxy, Ph-C₀₋₆alkoxy, Het-C₀₋₆alkoxy,OH, (CH₂)₁₋₆NR⁵⁸R⁵⁹, O(CH₂)₁₋₆NR⁵⁸R⁵⁹;

R⁵⁸, R⁵⁹=H, C₁₋₆alkyl, Ar—C₀₋₆alkyl; Het-C₀₋₆alkyl,

are more preferred embodiments of the present invention.

The following compounds of Formula IX are particularly preferred:

2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-(4-phenoxyphenylsulfonyl)]carbohydrazide;

2-[N-(N-benzyloxycarbonyl-L-alanyl)]-2′-[N-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide;

2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-(4-phenylbenzoyl)]carbohydrazide;

2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-(4-methoxybenzoyl)]carbohydrazide;

2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-(4-phenoxybenzoyl)]carbohydrazide;

2-(N-acetyl)-2′-[N′-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide;

2-[N-(N-acetyl-L-leucinyl)]-2′-[N′-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide;

2-[N-(N-acetyl-L-alanyl)]-2′-[N′-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide;

2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-[4-(N,N-dimethylaminomethyl)benzoyl)]]carbohydrazide;

2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-[4-hydroxy-[3-(4-morpholinomethyl)]]benzoyl]carbohydrazide;

2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-[4-(N,N-dimethylaminomethyl)benzyloxy]carbonyl-L-leucinyl]carbohydrazide;

2-(N-benzoyl)-2′-[N′-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide;

2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-[3-(4-morpholinomethyl)benzoyl]]carbohydrazide;

2-[N-(3-benzyloxybenzoyl)]-2′-[N′-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide;

2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-[4-[3-(N-N-dimethylamino)-1-propyloxy]enzoyl]]carbohydrazide;

2-[N-(2-benzyloxybenzoyl)]-2′-[N′-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide;

2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-[3-(4-pyridylmethoxy)benzoyl]]carbohydrazide;

2-[N-(4-benzyloxybenzoyl)]-2′-[N′-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide;

2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-(3-benzyloxy-5-methoxy)benzoyl]carbohydrazide;

2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-(3-benzyloxy-4,5-dimethoxy)benzoyl]carbohydrazide;

2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-(3-benzyloxy-5-ethoxy)benzoyl]carbohydrazide;

2-[N-(N-benzyloxycarbonylglycinyl)]-2′-[N′-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide;

2-[N-(3-benzyloxybenzoyl)]-2′-[N′-(N-benzyloxycarbonyl-L-prolinyl)]carbohydrazide;

2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-(4-phenylphenylacetyl)]carbohydrazide;

(2′S)-2-[N-(3-benzyloxybenzoyl)]-2′-[N′-(N-benzyloxycarbonyl-2-aminobutyryl)]carbohydrazide;

2,2′-[N,N′-[bis-(4-phenylphenylacetyl)]]carbohydrazide;

(2′RS)-2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[2-(4-phenylphenoxy)propionyl]carbohydrazide;

2-[N-(3-benzyloxybenzoyl)]-2′-[N′-(4-methylpentanoyl)]carbohydrazide;

(2RS,2′RS)-2,2′-[N,N′-[bis-[2-(4-phenylphenyl)₄methylpentanoyl)]]]carbohydrazide;

(2′RS)-2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-[2-(4-phenylphenyl)-4-methylpentanoyl)]]carbohydrazide;

(2′RS)-2-[N-(3-benzyloxybenzoyl)]-2′-[N′-[2-(4-phenylphenyl)₄-methylpentanoyl)]]carbohydrazide,

2-[N-(3-benzyloxybenzoyl)]-2′-[N′-(N-benzyloxycarbonyl-N-methyl-L-leucinyl)]carbohydrazide;

2-[N-(3-benzyloxybenzoyl)]-2′-[N′-[N-(2-pyridinylmethoxycarbonyl)-L-leucinyl]]carbohydrazide;

2-[N-[3-(4-pyridylmethoxy)benzoyl]]-2′-[N′-[N-(2-pyridinylmethoxycarbonyl)-L-leucinyl]]carbohydrazide;

(2RS)-2-[N-[2-(4phenylphenyl)₄methylpentanoyl)]]-2′-[N′-[N-(2-pyridinylmethoxycarbonyl)-L-leucinyl]]carbohydrazide;

2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-[2-(4-phenylphenyl)₄-methylpentanoyl)]]carbohydrazide;

2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-[2-(4-phenylphenyl)-4-methylpentanoyl)]]carbohydrazide;

2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-[N-(4-phenylphenyl)-N-(2-methylpropyl)carbamoyl]]carbohydrazide;

2-[N-(3-benzyloxybenzoyl)]-2′-[N′-(N-methyl-L-leucinyl)]carbohydrazide;

2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-(N-methyl-L-leucinyl)]carbohydrazide.

Compounds of Formula I wherein D=L—G—and Q=

are preferred embodiments of the present invention. For the sake ofconvenience, such compounds are referred to herein after as compounds ofFormula X.

With respect to Formula X:

More preferably G is

More preferably R⁶³ and R⁶⁴ are H and R⁶⁶ and R⁶⁹ are i-butyl.

More preferably R⁶⁵ is CH(R⁶⁹)NR⁶¹R⁷⁰, in which R⁶⁹ is i-butyl and R⁶¹is H. More preferably R⁷⁰ is R⁶²OC(O), in which R⁶² is

Alternately, R⁶⁵ is Ar or CH(R⁶⁹)Ar, in which Ar in said R⁶⁵ group is

More preferably, L is

CH(R⁶⁶)NR⁶⁰R⁶⁸, CH(R⁶⁶)Ar, NR⁶⁶R⁶⁷, CH(R⁶⁶)OAr′, Ar, or Het, in whichR⁶⁶ is i-butyl and Ar in said L group is

or Het in said L group is

More preferably L is NR⁶⁶R⁶⁷ or CH(R⁶⁶)R⁶⁰R⁶⁸.

One particularly preferred embodiment is a compound of Formula G:

Another particularly preferred embodiment is a compound of Formula H:

The following compounds of Formula X are most particularly preferred:

(1S)-N-[2-[(1-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-(4-phenoxyphenylsulfonyl)hydrazide;

(1S)-N-[4-[1-(N-benzyloxycarbonyl-L-leucinylamino)-3-methylbutyl]thiazol-2-ylcarbonyl]-N′-(N-benzyloxycarbonyl-L-leucinyl)hydrazide;

(1S)-N-[2-[(1-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-(4-phenylphenylacetyl)hydrazide;

(1S)-N-[2-[(1-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[3-(4-pryidinylmethoxy)benzoyl]hydrazide;

N-[2-(2-chlorophenoxymethyl)thiazolylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide;

N-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]-N′-[2-[4-(1,2,3-thiadiazol-4-yl)phenyl]thiazol-4-ylcarbonyl]hydrazide;

N-[2-[3-(4-chlorophenylsulfonylmethyl)thien-2-yl]thiazol-4ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide;

(1S,2′RS)-N-[2-[(1-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[2′-(4-phenylphenylacetyl)₄methylpentanoyl]hydrazide;

N-[2-(3-benzyloxyphenyl)thiazol-4-ylcarbonyl]-N′-[N-(2-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide;

(1RS)-N-[2-[1-(4-phenylphenyl)-3-methylbutyl]thiazolylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide;

N-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide;

N-[2-[N-methyl-N-(4-phenylphenyl)amino]thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide;

N-(N-benzyloxycarbonyl-L-leucinyl)-N′-[2-(4-phenylbenzyl)thiazol-4-ylcarbonyl]hydrazide;

N-[2-(4-phenylphenylbenzyl)thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)L-leucinyl]hydrazide;

N-(N-benzyloxycarbonyl-L-leucinyl)-N′-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]hydrazide;

N-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide;

N-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]-N′-[N-(3-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide;

N-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]-N′-[N-(2-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide;

N-(N-benzyloxycarbonyl-N-methyl-L-leucinyl)-N′-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]hydrazide;

N-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4ylcarbonyl]-N′-[N-(2-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide;

N-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]-N′-[N-(3-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide;and

N-[2-(2-methoxyphenyl)thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide.

Definitions

The present invention includes all hydrates, solvates, complexes andprodrugs of the compounds of this invention. Prodrugs are any covalentlybonded compounds which release the active parent drug according toFormula I in vivo. If a chiral center or another form of an isomericcenter is present in a compound of the present invention, all forms ofsuch isomer or isomers, including enantiomers and diastereomers, areintended to be covered herein. Inventive compounds containing a chiralcenter may be used as a racemic mixture, an enantiomerically enrichedmixture, or the racemic mixture may be separated using well-knowntechniques and an individual enantiomer may be used alone. In cases inwhich compounds have unsaturated carbon—carbon double bonds, both thecis (Z) and trans (E) isomers are within the scope of this invention. Incases wherein compounds may exist in tautomeric forms, such as keto-enoltautomers, each tautomeric form is contemplated as being included withinthis invention whether existing in equilibrium or predominantly in oneform.

The meaning of any substituent at any one occurrence in Formula I or anysubformula thereof is independent of its meaning, or any othersubstituent's meaning, at any other occurrence, unless specifiedotherwise.

Abbreviations and symbols commonly used in the peptide and chemical artsare used herein to describe the compounds of the present invention. Ingeneral, the amino acid abbreviations follow the IUPAC-IUB JointCommission on Biochemical Nomenclature as described in Eur. J. Biochem.,158, 9 (1984). The term “amino acid” as used herein refers to the D- orL-isomers of alanine, arginine, asparagine, aspartic acid, cysteine,glutamine, glutamnic acid, glycine, histidine, isoleucine, leucine,lysine, methionine, phenylalanine, proline, serine, threonine,tryptophan, tyrosine and valine.

“C₁₋₆alkyl” as applied herein is meant to include substituted andunsubstituted methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl andt-butyl, pentyl, n-pentyl, isopentyl, neopentyl and hexyl and the simplealiphatic isomers thereof. Any C₁₋₆alkyl group may be optionallysubstituted independently by one or two halogens, SR′, OR′, N(R′)₂,C(O)N(R′)₂, carbamyl or C₁₋₄alkyl, where R′ is C₁₋₆alkyl. C₀alkyl meansthat no alkyl group is present in the moiety. Thus, Ar—C₀alkyl isequivalent to Ar.

“C₃₋₁₁cycloalkyl” as applied herein is meant to include substituted andunsubstituted cyclopropane, cyclobutane, cyclopentane, cyclohexane,cycloheptane, cyclooctane, cyclononane, cyclodecane, cycloundecane.

“C₂₋₆ alkenyl” as applied herein means an alkyl group of 2 to 6 carbonswherein a carbon—carbon single bond is replaced by a carbon—carbondouble bond. C₂₋₆alkenyl includes ethylene, 1-propene, 2-propene,1-butene, 2-butene, isobutene and the several isomeric pentenes andhexenes. Both cis and trans isomers are included.

“C₂₋₆alkynyl” means an alkyl group of 2 to 6 carbons wherein onecarbon—carbon single bond is replaced by a carbon—carbon triple bond.C₂₋₆ alkynyl includes acetylene, 1-propyne, 2-propyne, 1-butyne,2-butyne, 3-butyne and the simple isomers of pentyne and hexyne.

“Halogen” means F, Cl, Br, and I.

“Ar” or “aryl” means=phenyl or naphthyl, optionally substituted by oneor more of Ph-C₀₋₆alkyl, Het-C₀₋₆alkyl, C₁₋₆alkoxy, Ph-C₀₋₆alkoxy,Het-Co -₆alkoxy, OH, (CH₂)₁₋₆NR⁵⁸R⁵⁹, O(CH₂)₁₋₆NR⁵⁸R⁵⁹; where R⁵⁸,R⁵⁹=H, C₁₋₆alkyl; Het-C₀₋₆alkyl, from C₁₋₄alkyl, OR′, N(R′)₂, SR′, CF₃,NO₂, CN, CO₂R′, CON(R′), F, Cl, Br and I.

As used herein “Het” or “heterocyclic” represents a stable 5- to7-membered monocyclic or a stable 7- to 10-membered bicyclicheterocyclic ring, which is either saturated or unsaturated, and whichconsists of carbon atoms and from one to three heteroatoms selected fromthe group consisting of N, O and S, and wherein the nitrogen and sulfurheteroatoms may optionally be oxidized, and the nitrogen heteroatom mayoptionally be quaternized, and including any bicyclic group in which anyof the above-defined heterocyclic rings is fused to a benzene ring. Theheterocyclic ring may be attached at any heteroatom or carbon atom whichresults in the creation of a stable structure, and may optionally besubstituted with one or two moieties selected from C₁₋₄alkyl, OR′,N(R′)₂, SR′, CF₃, NO₂, CN, CO₂R′, CON(R′), F, Cl, Br and I, where R′ isC₁₋₆alkyl. Examples of such heterocycles include piperidinyl,piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolodinyl,2-oxoazepinyl, azepinyl, pyrrolyl, 4-piperidonyl, pyrrolidinyl,pyrazolyl, pyrazolidinyl, imidazolyl, pyridyl, pyrazinyl, oxazolidinyl,oxazolinyl, oxazolyl, isoxazolyl, morpholinyl, thiazolidinyl,thiazolinyl, thiazolyl, quinuclidinyl, indolyl, quinolinyl,isoquinolinyl, benzimidazolyl, benzopyranyl, benzoxazolyl, furyl,pyranyl, tetrahydrofuryl, tetrahydropyranyl, thienyl, benzoxazolyl,thiamorpholinyl sulfoxide, thiamorpholinyl sulfone, and oxadiazolyl.

“HetAr” or “heteroaryl” means any heterocyclic moiety encompassed by theabove definition of Het which is aromatic in character, e.g., pyridine.

It will be appreciated that the heterocyclic ring described when N=

includes thiazoles, oxazoles, triazoles, thiadiazoles, oxadiazoles,isoxazoles, isothiazols, imidazoles, pyrazines, pyridazines,pyrimidines, triazines and tetrazines which are available by routinechemical synthesis and are stable. The single and double bonds (i.e., )in such heterocycles are arranged based upon the heteroatoms present sothat the heterocycle is aromatic (e.g., it is a heteroaryl group). Theterm heteroatom as applied herein refers to oxygen, nitrogen and sulfur.When the heteroaryl group comprises a five membered ring, W ispreferably an electron withdrawing group, such as halogen, —CN, —CF₃,—NO₂, —COR⁷, —CO₂R⁶, —CONHR⁶, —SO₂NHR⁶, —NHSO₂R⁶, —NHCOR⁷, —O—COR⁶, —SR⁶or NR′R⁶, or a similar electron withdrawing substituent as known in theart.

Certain radical groups are abbreviated herein. t-Bu refers to thetertiary butyl radical, Boc refers to the t-butyloxycarbonyl radical,Fmoc refers to the fluorenylmethoxycarbonyl radical, Ph refers to thephenyl radical, Cbz refers to the benzyloxycarbonyl radical.

Certain reagents are abbreviated herein. DCC refers todicyclohexylcarbodiimide, DMAP is 2,6-dimethylaminopyridine, EDC refersto N-ethyl-N′(dimethylaminopropyl)-carbodiimide. HOBT refers to1-hydroxybenzotriazole, DMF refers to dimethyl formamide, BOP refers tobenzotriazol-1-yloxy-tris(dimethylamino)phosphonium hexafluorophosphate,DMAP is dimethylaminopyridine, Lawesson's reagent is2,4-bis(4-methoxyphenyl)-1,3-dithia-2,4-diphosphetane-2,4-disulfide, NMMis N-methylmorpholine, TFA refers to trifluoroacetic acid, TFAA refersto trifluoroacetic anhydride and THF refers to tetrahydrofuran. Jonesreagent is a solution of chromium trioxide, water, and sulfuric acidwell-known in the art.

Methods of Preparation

Compounds of Formula II wherein X=CH, Y=S and Z=N, and W=CO₂R⁷ CN, orCONR′R⁷ may be conveniently prepared by methods analogous to thosedescribed in Scheme 1.

1-Scheme 1 is treated with isobutyl chloroformate and N-methylmorpholinein ethyl acetate to give a mixed anhydride which is treated withdiazomethane in ether to provide 2-Scheme 1. The diazoketone ishalogenated using 30% HBr in acetic acid in ethyl acetate/ether solutionto provide 3-Scheme 1. This material is treated with ethyl thiooxamatein refluxing ethanol to give 4-Scheme 1. The thiazole carboxylic esteris saponified by treatment with a hydroxide base (such as potassiumhydroxide, sodium hydroxide or lithium hydroxide) to yield carboxylicacid 5-Scheme 1. The carboxylic acid is treated with isobutylchloroformate and N-methylmorpholine, followed by gaseous ammonia toprovide primary amide 6-Scheme 1 (R³═H). The primary amide is treatedwith TFAA and pyridine in dichloromethane to provide 7-Scheme 1.Alternatively, 5-Scheme 1 can be converted to substituted amides,6-Scheme 1, by treatment with alkyl amines (such as benzylamine,2-phenylethylamine or isobutylamine) and a peptide coupling reagent(such as BOP, EDC.HCl/1-HOBT or N-methylmorpholine/isobutylchloroformate) in an aprotic solvent (such as dichloromethane or DMF).The carboxylic acid 5-Scheme 1 can be converted to carboxylic esters8-Scheme 1 by treatment with a primary or secondary alcohol (such as2,2,2-trifluoroethanol, isobutyl alcohol, benzyl alcohol or phenol) anda dehydrating reagent (such as DCC/DMAP, EDCI or Boc₂O/pyridine) in anaprotic solvent (such as dichloromethane or ether). WhenR²=9-fluorenylmethoxy, treatment of 4-Scheme 1 with piperidine in DMFgives 9-Scheme 1. Treatment of 9-Scheme 1 with a carboxylic acid (suchas N-Cbz-L-phenylalanine or N-Cbz-L-leucinyl-L-leucine) and a peptidecoupling reagent (such as BOP) in an aprotic solvent (such asdichloromethane) provides 10-Scheme 1.

Compounds of Formula II wherein X=CH, Y=S and Z=N are prepared bymethods analogous to those described in Scheme 1A. 1-Scheme 1A istreated with iodomethane in an aprotic solvent (such as THF) to afford2-Scheme 1A, which is treated with a primary amine in a protic solvent(such as isopropanol) to give 3-Scheme 1A. this material is then treatedwith a bromomethyl ketone in a protic solvent (such as ethanol) toprovide 4-Scheme 1A.

Compounds of Formula II wherein X=S, Y=CH and Z=N may be convenientlyprepared by methods analogous to those described in Scheme 2. 1-Scheme 2is treated with isobutyl chloroformate, N-methylmorpholine and ammoniain THF to provide 2-Scheme 2. This material is converted to thethioamide, 3-Scheme 2, by treatment with Lawesson's reagent in anaprotic solvent (such as THF or toluene). 3-Scheme 2 is converted to thethiazole by condensation with a α-ketoester bearing a suitable leavinggroup for displacement by a sulfur nucleophile (Cl, Br, I, OMs, O-p-Tos)in dichloromethane. 4-Scheme 2 is treated with TFA to provide 5-Scheme2. This material is treated with a carboxylic acid (such asN-Cbz-L-leucine, N-Cbz-D-leucine or N-Cbz-L-leucinyl-L-leucine) and apeptide coupling reagent (such as BOP, EDC.HCl/1-HOBT orN-methylmorpholine/isobutyl chloroformate) in an aprotic solvent (suchas dichloromethane, DMF or THF) to yield 6-Scheme 2. This material issaponified by treatment with a hydroxide base (such as potassiumhydroxide, sodium hydroxide or lithium hydroxide) to yield carboxylicacid 7-Scheme 2.

Compounds of Formula II wherein X=S, Y=CH and Z=N may also be preparedby methods analogous to those described in Scheme 2A. 1-Scheme 2A istreated with a tert-butoxycarbonyl-protected amino acid (such asN-tert-butoxycarbonyl-L-leucine) and a peptide coupling reagent (such asBOP, EDC.HCl/1-HOBT or N-methylmorpholine/isobutyl chloroformate) in anaprotic solvent (such as dichloromethane, DMF or THF) to yield 2-Scheme2A, which is treated with trifluoroacetic acid to provide 3-Scheme 2A.This material is treated with a chloroformate (such as 2-biphenylmethylchloroformate, 2-benzylbenzyl chloroformate, 2-naphthylmethylchloroformate or 2-phenoxybenzyl chloroformate) and a tertiary aminebase (such as diisopropylethylamine) in an aprotic solvent (such asmethylene chloride) to provide 4-Scheme 2A.

Compounds of Formula II wherein X and Y=N, and Z=S may be convenientlyprepared by methods analogous to those described in Scheme 3. 1-Scheme 3is treated with di-tert-butyl dicarbonate and triethylamine in THF toprovide 2-Scheme 3. This material is treated with hydrazine hydrate inmethanol to provide 3-Scheme 3. The hydrazide is acylated by treatmentwith ethyl oxalyl chloride and pyridine in dichloromethane to provide4-Scheme 3. This material is converted to the thiadiazole, 5-Scheme 3,by treatment with Lawesson's reagent in an aprotic solvent (such as THFor toluene). 5-Scheme 3 is treated with TFA to provide 6-Scheme 3. Thismaterial is treated with a carboxylic acid (such as N-Cbz-L-leucine) anda peptide coupling reagent (such as BOP, EDC.HCl/1-HOBT orN-methylmorpholine/isobutyl chloroformate) in an aprotic solvent (suchas dichloromethane, DMF or THF) to yield 7-Scheme 3.

Compounds of Formula II wherein X and Y=N, and Z=O, and W=CO₂Et or CONH₂may be conveniently prepared by methods analogous to those described inScheme 4. 1-Scheme 4 is treated with thionyl chloride and pyridine inether, followed by refluxing in toluene to provide 2-Scheme 4. Theresultant oxadiazole is treated with TFA to provide 3-Scheme 4. Thismaterial is treated with a carboxylic acid (such as N-Cbz-L-leucine) anda peptide coupling reagent (such as BOP, EDC.HCl/1-HOBT orN-methylmorpholine/isobutyl chloroformate) in an aprotic solvent (suchas dichloromethane, DMF or THF) to yield 4-Scheme 4. The carboxylicester is treated with ammonia in methanol to yield 5-Scheme 4.

Compounds of Formula II wherein X and Y=N, and Z=O, and W=SH may beconveniently prepared by methods analogous to those described in Scheme5. 1-Scheme 5 and 2-Scheme 5 are treated with a peptide coupling reagent(such as BOP, EDC.HCl/1-HOBT or N-methylmorpholine/isobutylchloroformate) in an aprotic solvent (such as dichloromethane, DMF orTHF) to yield 3-Scheme 5. This material is treated with hydrazinehydrate in methanol to provide 4-Scheme 5. Treatment of 4-Scheme 5 withthiophosgene and triethylamine in chloroform provides 5-Scheme 5.

Compounds of Formula II wherein X=CH, Y=S and Z=N, and W=SH or NH₂ maybe conveniently prepared by methods analogous to those described inScheme 6. Condensation of 1-Scheme 6 with ammonium dithiocarbamate inethanol yielded 2-Scheme 6. Alternatively, 1-Scheme 6 can be condensedwith thiourea in ethanol to give 3-Scheme 6.

Compounds of Formula II wherein X=CH, Y=N and Z=N and W=C may beprepared by methods analogous to those described in Scheme 7. Treatmentof 1-Scheme 7 with diethylamine N-oxide should provide 2-Scheme 7.Condensation of 2-Scheme 7 with a 2,3-diaminocarboxylic acid should thenprovide 3-Scheme 7, which may be converted to a variety of carboxylicacid derivatives using procedures previously described in other schemes.

Compounds of Formula III may be generally prepared by methods common inthe art of organic chemistry for coupling carboxylic acid derivatives tohydrazine. Schemes 8, 9 and 10 are illustrative of a method to preparecompounds wherein B or E is a heterocycle. Compounds of Formula X may beconveniently prepared by methods analogous to those described in Schemes8, 9 and 19-23.

Compounds wherein X=CH, Y=S and Z=N, are prepared by methods analogousto those described in Scheme 22. 1-Scheme 8 is treated with isobutylchloroformate and N-methylmorpholine in ether to give a mixed anhydridewhich is treated with diazomethane in ether to provide 2-Scheme 8. Thediazoketone is halogenated using 30% HBr in acetic acid in ethersolution to provide 3-Scheme 8. This material is treated with ethylthiooxamate in refluxing ethanol to give 4-Scheme 8. The thiazolecarboxylic ester is treated with a hydrazine (such as hydrazinemonohydrate or methyl hydrazine) in ethanol to yield 5-Scheme 8. Thismaterial is treated with a carboxylic acid (such as N-Cbz-L-leucine) anda peptide coupling reagent (such as EDC.HCl/1-HOBT) in an aproticsolvent (such as DMF) to provide 6-Scheme 8.

Compounds wherein X=S, Y=CH and Z=N, are prepared by methods analogousto those described in Scheme 9.

1-Scheme 9 is converted to 2-Scheme 9 by treatment with isobutylchloroformate, N-methylmorpholine and ammonia in THF. 2-Scheme 9 istreated with Lawesson's reagent in THF to provide the thioamide 3-Scheme9. This material is converted to the thiazole by condensation with anα-ketoester followed by treatment with trifluoroacetic anhydride andpyridine in methylene chloride to afford 4-Scheme 21 which is convertedto 5-Scheme 9 by treatment with hydrazine monohydrate. This material istreated with a sulfonyl chloride (such as 4-phenoxybenzenesulfonylchloride) and pyridine in an aprotic solvent (such as dichloromethane)to provide 6-Scheme 9. Alternatively, 6-Scheme 9 may be prepared bytreatment with a carboxylic acid (such as N-benzyloxycarbonyl-L-leucine,N-benzyloxycarbonyl-N-methyl-L-leucine,N-(2-pyridinylmethoxycarbonyl)-L-leucine,N-(3-pyridinylmethoxycarbonyl)-L-leucine,N-(4-pyridinylmethoxycarbonyl)-L-leucine, 4-biphenylacetic acid,3-(4-pyridinylmethoxy)benzioc acid, or4-methyl-2-(4-phenylphenyl)pentanoic acid) and a peptide couplingreagent (such as EDC.HCl/1-HOBT) in an aprotic solvent (such as DMF).

Compounds wherein B=

are prepared by routine methods of peptide synthesis as illustrated forinstance by Scheme 10.

Treatment of a mixture of 1-Scheme 10 and 2-Scheme 10 with a peptidecoupling reagent (such as BOP or EDC.HCl/1-HOBT) in an aprotic solvent(such as DMF or dichloromethane) provides 3-Scheme 10. This material istreated with hydrazine hydrate in ethanol to yield 4-Scheme 10, which istreated with a carboxylic acid (such as N-Cbz-L-leucine) and a peptidecoupling reagent (such as BOP or EDC.HCl/1-HOBT) in an aprotic solvent(such as DMF or dichloromethane) to provide 5-Scheme 10.

Compounds of Formula IV wherein R²², R²³, R²⁴ are H, and R²¹=R²⁶ areprepared by methods analogous to those described in Scheme 11.

Symmetric compounds of the Formula IX having RCO as the terminalsubstituent on both sides are prepared by methods analogous to thosedescribed in Scheme 11. Treatment of 1-Scheme 11 with a carboxylic acid(such as 4-biphenylacetic acid or 4-methyl-2-(4-phenylphenyl)pentanoicacid) and a peptide coupling reagent (such as EDC.HCl/1-HOBT) in anaprotic solvent (such as DMF) provides 2-Scheme 11.

Nonsymmnetric compounds of the Formula IX, and compounds of Formula IVwherein R²², R²³, R²⁴ and R²⁵ are H, and R²¹≠R²⁶, are prepared bymethods analogous to those described in Scheme 12.

Treatment of 1-Scheme 12 with hydrazine hydrate in a protic solvent(such as methanol or ethanol) provides 2-Scheme 12, which is treatedphosgene in toluene to afford 3-Scheme 12. This material is treated withhydrazine hydrate in a protic solvent (such as methanol or ethanol) toprovide 4-Scheme 12. Treatment of 4-Scheme 12 with a sulfonyl chloride(such as 4-phenoxyphenylsulfonyl chloride), an acid chloride (such asbenzoyl chloride), or a carbamoyl chloride (such asN-(2-methylpropyl)-N-(4-phenylphenyl)carbamoyl chloride) and pyridine inDMF affords 5-Scheme-12. Alternatively, 5-Scheme-12 may be prepared bytreatment of 4-Scheme 12 with a carboxylic acid (such asN-benzyloxycarbonyl-L-alanine, N-benzyloxycarbonyl-L-proline,N-benzyloxycarbonylglycine, (S)-N-benzyloxycarbonyl-2-aminobutyric acid,N-benzyloxycarbonyl-N-methyl-L-leucine,N-tert-butoxycarbonyl-N-methyl-L-leucine, N-acetyl-L-leucine,N-acetyl-L-alanine, N-(2-pyridinylmethoxycarbonyl)-L-leucine,N-[4-(N,N-dimethylaminomethyl)benzyloxycarbonyl]-L-leucine,4-phenylbenzoic acid, 4-methoxybenzioc acid, 4-phenoxybenzoic acid,4-(N,N-dimethylanunomethyl)benzoic acid,4-hycroxy-3-[N-(4-morpholinomethyl)]benzoic acid,3-[N-(4-morpholinomethyl)]benzoic acid, 2-benzyloxybenzoic acid,3-benzyloxybenzoic acid, 4-benzyloxybenzoic acid,4-(3-dimethylaminomethylpropoxy)benzoic acid,3-benzyloxy-5-methoxybenzoic acid, 3-benzyloxy-4,5-dimethoxybenzoicacid, 3-benzyloxy-5-ethoxybenzoic acid, 3-(4-pyridinylmethoxy)benzoicacid, 4-biphenylacetic acid, 2-(4-phenylphenoxy)propionic acid or4-methyl-2-(4-phenylphenyl)pentanoic acid) and a peptide couplingreagent (such as BOP, EDC.HCl/1-HOBT or N-methylmorpholine/isobutylchioroformate) in an aprotic solvent (such as dichloromethane, DMF orTHF). 5-Scheme-12 may also be prepared by treatment of 4-Scheme 12 withan anhydride (such as acetic anhydride). Alternatively, 3 Scheme 12 maybe converted directly to 5-Scheme-I by treatment with a hydrazide (suchas 4-methylpentanoyl hydrazide orN-methyl-N-benzyloxycarbonyl-L-leucinyl hydrazide).

Nonsymmetric compounds of Formula IV, wherein R²³≠H are prepared bymethods analogous to those described in Scheme 12A. 1-Scheme 12A istreated with an aldehyde (such as benzaldehyde) in a protic solvent(such as ethanol) and the resulting imine is treated with borane-THFcomplex to afford 2-Scheme 12A, which is subsequently treated withphosgene in toluene to afford 3-Scheme 12A. This material is treatedwith hydrazine hydrate in a protic solvent (such as methanol or ethanol)to provide 4-Scheme 12A. Treatment of 4-Scheme 12A with a carboxylicacid (such as N-benzyloxycarbonyl-L-leucine) and a peptide couplingreagent (such as BOP, EDC.HCl/1-HOBT or N-methylmorpholine/isobutylchloroformate) in an aprotic solvent (such as dichloromethane, DMF orTHF) to yield 5-Scheme-12A.

Compounds of Formulae V-VII may be conveniently prepared by methodsanalogous to those described in Schemes 13-16.

1,3-Bis-amido propan-2-ones may be prepared by acylation of1,3-diamino-propan-2-ol 1-Scheme 13 with a carboxylic acid 2-Scheme 13or a mixture of 2 different carboxylic acids (2 and 3) in equimolaramounts and a coupling reagent such as a dialkyl carbodiimide such asDCC or EDCI or HBTU/N-methyl morpholine, followed by oxidation of thecarbinol to a ketone with an oxidant such as Jones reagent.

1,3-Bis-sulfonamido propanones may be prepared by sulfonylation of1,3-diamino-propan-2-ol 1-Scheme 14 with a sulfonyl chloride 2-Scheme 14and a base such as N-methyl morpholine, followed by oxidation of thecarbinol to a ketone with an oxidant such as Jones reagent.

1-Amido-3-sulfonamido propanones may be prepared by acylation of1,3-diamino-propan-2-ol 1-Scheme 15 with a carboxylic acid 2-Scheme 15and a coupling reagent such as a carbodiimide or HBTU/N-methylmorpholine, followed by treatment with an appropriate sulfonyl chloride3-Scheme 15 and a base such as N-methyl morpholine, followed byoxidation of the carbinol to a ketone with an oxidant such as Jonesreagent.

1-Amido-3-sulfonamido alkan-2-ones that are larger than propan-2-one,such as butan-2-one or 5-methyl-hexan-2-one, can be prepared byconverting an N-protected peptide such as Cbz-leu-leu-OH 1-Scheme 16 toits bromo methyl ketone 3-Scheme 16 via a diazo methyl ketone 2-Scheme16. Then, the bromide 3-Scheme 1 is displaced with sodium azide to givethe corresponding azide 4-Scheme 16. Reduction of the carbonyl with areducing agent such as sodium borohydride gives alcohol 5-Scheme 16.Subsequent reduction of the azide with a reducing agent such as1,3-propandithiol gives the free amine 6-Scheme 16. Acylation orsulfonylation of the amine gives amide or sulfonamide 7-Scheme 16.Finally, oxidation of the carbinol with an oxidant such as Jones givesthe desired compounds.

Compounds of Formula VIII may be conveniently made using methodsanalogous to those in Schemes 17 and 18.

Azide opening of glycidol 1-Scheme 17, followed by tosylation of theprimary alcohol gave tosylate 2-Scheme 17, which was coupled to Ellmanpolymer 3-Scheme 17 as described by described in J. Med. Chem. 1995, 38,1427-1430 to produce polymer 4-Scheme 17, which was reacted with benzylamine in toluene, then washed extensively with various solvents. Then,the azide was reduced with 1,3-propanedithiol in MeOH, triethylamine,then was washed extensively with various solvents. Coupling ofCbz-leucine 6-Scheme 17 with the diamine 5-Scheme 1l with equimolaramounts and a coupling reagent such as a dialkyl carbodiimide such asDCC or EDCI or HBTU/N-methyl morpholine. Cleavage of the ether linkageto an alcohol was accomplished with trifluoroacetic acid with variousscavengers. Finally, oxidation of the carbinol to a ketone 7-Scheme 17with an oxidant such as Jones reagent provided the desired finalproduct.

N-(2,3-Epoxypropyl)phthalimide 1-Scheme 18 (Aldrich) was reluxed with anamine such as 4-pyridiyl methyl amine in isopropanol. The secondaryamine 2-Scheme 18 was then acylated with an acylating agent such as Cbzleucine or a sulfonylating reagent such as 2-dibenzofuransulfonylchloride and base such as N-methyl morpholine in DMF. The phthalimidewas then removed with hydrazine in MeOH and the resulting free amine wasacylated with an acylating agent such as Cbz leucine or a sulfonylatingreagent such as 2-dibenzofuransulfonyl chloride and base such asN-methyl morpholine in DMF.

Compounds of Formula IX may conveniently be made using methods analogousto those in Schemes 19 and 20.

Compounds of Formula X may be conveniently made using methods analogousto those described in Schemes 21-27.

Compounds wherein X=CH, Y=S, Z=N and R⁴≠H, are prepared by methodsanalogous to those described in Scheme 19. Carboxylic ester 1-Scheme 19is treated with a hydroxide base (such as lithoum hydroxide, sodiumhydroxide or potassium hydroxide) in methanol/water to provide 2-Scheme19. 3-Scheme 19 is treated with a hydrazine (such as methylhydrazine) ina protic solvent (such as ethanol) to give 4-Scheme 19. 2-Scheme 12 and4-Scheme 19 are coupled by treatment with a peptide coupling reagent(such as EDC.HCl/1-HOBT) in an aprotic solvent (such as DMF) to provide5-Scheme 19.

Compounds wherein X=S, Y=CH, Z=N and V=2-methoxyphenyl or2-benzyloxyphenyl, are prepared by methods analogous to those describedin Scheme 20. Ethyl bromopyruvate (1-Scheme 20) is treated with thioureain refluxing ethanol to provide 2-Scheme 20, which is treatedsuccessively with sodium nitrite and copper (I) bromide in 16% aqueousHBr, and the product was heated in ethanol with a catalytic amount ofHBr to give 3-Scheme 20. Treatment of this material with an arylboronicacid (such as 2-benzyloxyphenylboronic acid),tetrakis(triphenylphosphine)palladium(O) and cesium fluoride inrefluxing DME provides 4Scheme 20. Alternatively, 4Scheme 20, may beprepared by treatment of 3-Scheme 20 with an arylstannane (such as2-trimethylstannylanisole) and tetrakis(triphenylphosphine)palladium(O)in refluxing toluene. Treatment of 4 Scheme 20 with hydrazine hydrate inethanol provides 5-Scheme 20. which is treated with a carboxylic acid(such as N-benzyloxycarbonyl-N-methyl-L-leucine,N-(2-pyridinylmethoxycarbonyl)-L-leucine,N-(3-pyridinylmethoxycarbonyl)-L-leucine orN-(4-pyridinylmethoxycarbonyl)-L-leucine) and a peptide coupling reagent(such as EDC.HCl/1-HOBT) in an aprotic solvent (such as DMF) to provide6-Scheme 20.

Compounds wherein X=S, Y=CH, Z=N and V=NR⁶⁶R⁶⁷, are prepared by methodsanalogous to those described in Scheme 21. An acid chloride (1-Scheme21) is treated with a primary amine (such as 4-aminobiphenyl or aniline)and pyridine in an aprotic solvent (such as methylene chloride) toprovide 2-Scheme 21, which is treated with lithium aluminum hydride inTHF to afford 3-Scheme 25. Treatment of 3-Scheme 21 with thiophosgeneand pyridine in methylene chloride, followed by treatment with ammoniain methanol provides 4-Scheme 21. Alternatively, 4-Scheme 21 may beprepared by treatment of 3-Scheme 21 with benzoyl isothiocyanate,followed by treatment of the intermediate benzoyl thiourea withpotassium carbonate in methanol/water. 4-Scheme 21 is treated withhydrazine hydrate in ethanol to give 5-Scheme 21. Treatment of 5-Scheme21 with a carboxylic acid (such asN-(2-pyridinylmethoxycarbonyl)-L-leucine,N-(3-pyridinylmethoxycarbonyl)-L-leucine orN-(4pyridinylmethoxycarbonyl)-L-leucine) and a peptide coupling reagent(such as EDC.HCl/1-HOBT) in an aprotic solvent (such as DMF) affords6-Scheme 21.

Compounds wherein X and Y=N, and Z=NH, are prepared by methods analogousto those described in Scheme 26. 1-Scheme 22 is treated with hydrazinehydrate in ethanol to give 2-Scheme 22, which is heated with a mixedanhydride to provide triazole 3-Scheme 22. This material is treated withhydrazine hydrate to provide 4-Scheme 22, which is treated with acarboxylic acid (such as N-benzyloxycarbonyl-L-leucine) and a peptidecoupling reagent (such as EDC.HCl/1-HOBT) in an aprotic solvent (such asDMF) to provide 5-Scheme 22.

Compounds wherein X=S, Y=CH, Z=N, L=CH(R⁶⁶)NR⁶⁰R⁶⁸ where R⁶⁸. Boc orCbz, or R⁶⁵=CH(R⁶⁹)NR⁶¹R⁷⁰ where R⁷⁰≠Boc or Cbz are prepared by methodsanalogous to those described in Scheme 27. 1-Scheme 23 is treated withtrifluoroacetic acid to provide 2-Scheme 23. This material is treatedwith a carboxylic acid (such as pryazinecarboxylic acid, isonicotinicacid, 4-imidazolylacetic acid or pipecolic acid) and a peptide couplingreagent (such as EDC.HCl/1-HOBT) in an aprotic solvent (such as DMF) toprovide 3-Scheme 23. 3-Scheme 23 may also be prepared by treatment of2-Scheme 23 with a sulfonyl chloride (such as 2-pyridinesulfonylchloride) and a tertieary amine base (such as diisopropylethylamine) inan aprotic solvent (such as methylene chloride). Alternatively,treatment of 4-Scheme 23 with trifluoroacetic acid provides 5 Scheme 23.

1,3-Diamino-propan-2-ol (or an N-alkyl substituted diamino-propanol) iscoupled to a protected leucine analog (either Cbz- or Boc-) and anothercarboxylic acid or sulfonyl chloride. Removal of the protective group,followed by acylation or sulfonylation, and oxidation of the alcoholprovides the desired compounds.

N-Allyl amine (or a N-alkyl-N-allyl amine) is coupled to a Cbz-aminoacid (or sulfonylated with an aryl sulfonyl chloride), then the alkeneis epoxidized with a peracid (or dimethyl diooxirane). The epoxide isopened with a subsituted amine, then the amine is acylated orsulfonylated. Final oxidation gives the desired ketones.

The starting materials used herein are commercially available aminoacids or are prepared by routine methods well known to those of ordinaryskill in the art and can be found in standard reference books, such asthe COMPENDIUM OF ORGANIC SYNTHETIC METHODS, Vol. I-VI (published byWiley-Interscience).

Coupling methods to form amide bonds herein are generally well known tothe art. The methods of peptide synthesis generally set forth byBodansky et al., THE PRACTICE OF PEPTIDE SYNTHESIS, Springer-Verlag,Berlin, 1984; E. Gross and J. Meienhofer, THE PEPTIDES, Vol. 1, 1-284(1979); and J. M. Stewart and J. D. Young, SOLID PHASE PEPTIDESYNTHESIS, 2d Ed., Pierce Chemical Co., Rockford, Ill., 1984. aregenerally illustrative of the technique and are incorporated herein byreference.

Synthetic methods to prepare the compounds of this invention frequentlyemploy protective groups to mask a reactive functionality or minimizeunwanted side reactions. Such protective groups are described generallyin Green, T. W, PROTECTIVE GROUPS IN ORGANIC SYNTHESIS, John Wiley &Sons, New York (1981). The term “amino protecting groups” generallyrefers to the Boc, acetyl, benzoyl, Fmoc and Cbz groups and derivativesthereof as known to the art. Methods for protection and deprotection,and replacement of an amino protecting group with another moiety arewell known.

Acid addition salts of the compounds of Formula I are prepared in astandard manner in a suitable solvent from the parent compound and anexcess of an acid, such as hydrochloric, hydrobromic, hydrofluoric,sulfuric, phosphoric, acetic, trifluoroacetic, maleic, succinic ormethanesulfonic. Certain of the compounds form inner salts orzwitterions which may be acceptable. Cationic salts are prepared bytreating the parent compound with an excess of an alkaline reagent, suchas a hydroxide, carbonate or alkoxide, containing the appropriatecation; or with an appropriate organic amine. Cations such as Li⁺, Na⁺,K⁺, Ca⁺⁺, Mg⁺⁺ and NH₄ ⁺ are specific examples of cations present inpharmaceutically acceptable salts. Halides, sulfate, phosphate,alkanoates (such as acetate and trifluoroacetate), benzoates, andsulfonates (such as mesylate) are examples of anions present inpharmaceutically acceptable salts.

This invention also provides a pharmaceutical composition whichcomprises a compound according to Formula I and a pharmaceuticallyacceptable carrier, diluent or excipient. Accordingly, the compounds ofFormula I may be used in the manufacture of a medicament. Pharmaceuticalcompositions of the compounds of Formula I prepared as hereinbeforedescribed may be formulated as solutions or lyophilized powders forparenteral administration. Powders may be reconstituted by addition of asuitable diluent or other pharmaceutically acceptable carrier prior touse. The liquid formulation may be a buffered, isotonic, aqueoussolution. Examples of suitable diluents are normal isotonic salinesolution, standard 5% dextrose in water or buffered sodium or ammoniumacetate solution. Such formulation is especially suitable for parenteraladministration, but may also be used for oral administration orcontained in a metered dose inhaler or nebulizer for insufflation. Itmay be desirable to add excipients such as polyvinylpyrrolidone,gelatin, hydroxy cellulose, acacia, polyethylene glycol, mannitol,sodium chloride or sodium citrate.

Alternately, these compounds may be encapsulated, tableted or preparedin an emulsion or syrup for oral administration. Pharmaceuticallyacceptable solid or liquid carriers may be added to enhance or stabilizethe composition, or to facilitate preparation of the composition. Solidcarriers include starch, lactose, calcium sulfate dihydrate, terra alba,magnesium stearate or stearic acid, talc, pectin, acacia, agar orgelatin. Liquid carriers include syrup, peanut oil, olive oil, salineand water. The carrier may also include a sustained release materialsuch as glyceryl monostearate or glyceryl distearate, alone or with awax. The amount of solid carrier varies but, preferably, will be betweenabout 20 mg to about 1 g per dosage unit. The pharmaceuticalpreparations are made following the conventional techniques of pharmacyinvolving milling, mixing, granulating, and compressing, when necessary,for tablet forms; or milling, mixing and filling for hard gelatincapsule forms. When a liquid carrier is used, the preparation will be inthe form of a syrup, elixir, emulsion or an aqueous or non-aqueoussuspension. Such a liquid formulation may be administered directly p.o.or filled into a soft gelatin capsule.

For rectal administration, the compounds of this invention may also becombined with excipients such as cocoa butter, glycerin, gelatin orpolyethylene glycols and molded into a suppository.

Utility of the Present Invention

The compounds of Formula I are useful as protease inhibitors,particularly as inhibitors of cysteine and serine proteases, moreparticularly as inhibitors of cysteine proteases, even more particularlyas inhibitors of cysteine proteases of the papain superfanmily, yet moreparticularly as inhibitors of cysteine proteases of the cathepsinfamily, most particularly as inhibitors of cathepsin K. The presentinvention also provides useful compositions and formulations of saidcompounds, including pharmaceutical compositions and formulations ofsaid compounds.

The present compounds are useful for treating diseases in which cysteineproteases are implicated, including infections by pneumocystis care,trypsanoma cruzi, trypsanoma brucei, and Crithidia fusiculata; as wellas in schistosomiasis, malaria, tumor metastasis, metachromaticleukodystrophy, muscular dystrophy, amytrophy; and especially diseasesin which cathepsin K is implicated, most particularly diseases ofexcessive bone or cartilage loss, including osteoporosis, gingivaldisease including gingivitis and periodontitis, arthritis, morespecifically, osteoarthritis and rheumatoid arthritis, Paget's disease;hypercalcemia of malignancy, and metabolic bone disease.

Metastatic neoplastic cells also typically express high levels ofproteolytic enzymes that degrade the surrounding matrix, and certaintumors and metastatic neoplasias may be effectively treated with thecompounds of this invention.

The present invention also provides methods of treatment of diseasescaused by pathological levels of proteases, particularly cysteine andserine proteases, more particularly cysteine proteases, even moreparticularly as inhibitors of cysteine proteases of the papainsuperfamily, yet more particularly cysteine proteases of the cathepsinfamily, which methods comprise administering to an animal, particularlya mammal, most particularly a human in need thereof a compound of thepresent invention. The present invention especially provides methods oftreatment of diseases caused by pathological levels of cathepsin K,which methods comprise administering to an animal, particularly amammal, most particularly a human in need thereof an inhibitor ofcathepsin K, including a compound of the present invention. The presentinvention particularly provides methods for treating diseases in whichcysteine proteases are implicated, including infections by pneumocystiscarinii, trypsanoma cruzi, trypsanoma brucei, and Crithidia fusiculata;as well as in schistosomiasis, malaria, tumor metastasis, metachromaticleukodystrophy, muscular dystrophy, amytrophy, and especially diseasesin which cathepsin K is implicated, most particularly diseases ofexcessive bone or cartilage loss, including osteoporosis-, gingivaldisease including gingivitis and periodontitis, arthritis, morespecifically, osteoarthritis and rheumatoid arthritis, Paget's disease,hypercalcemia of malignancy, and metabolic bone disease.

This invention further provides a method for treating osteoporosis orinhibiting bone loss which comprises internal administration to apatient of an effective amount of a compound of Formula I, alone or incombination with other inhibitors of bone resorption, such asbisphosphonates (i.e., allendronate), hormone replacement therapy,anti-estrogens, or calcitonin. In addition, treatment with a compound ofthis invention and an anabolic agent, such as bone morphogenic protein,iproflavone, may be used to prevent bone loss or to increase bone mass.

For acute therapy, parenteral administration of a compound of Formula Iis preferred. An intravenous infusion of the compound in 5% dextrose inwater or normal saline, or a similar formulation with suitableexcipients, is most effective, although an intramuscular bolus injectionis also useful. Typically, the parenteral dose will be about 0.01 toabout 100 mg/kg; preferably between 0.1 and 20 mg/kg, in a manner tomaintain the concentration of drug in the plasma at a concentrationeffective to inhibit cathepsin K. The compounds are administered one tofour times daily at a level to achieve a total daily dose of about 0.4to about 400 mg/kg/day. The precise amount of an inventive compoundwhich is therapeutically effective, and the route by which such compoundis best administered, is readily determined by one of ordinary skill inthe art by comparing the blood level of the agent to the concentrationrequired to have a therapeutic effect.

The compounds of this invention may also be administered orally to thepatient, in a manner such that the concentration of drug is sufficientto inhibit bone resorption or to achieve any other therapeuticindication as disclosed herein. Typically, a pharmaceutical compositioncontaining the compound is administered at an oral dose of between about0.1 to about 50 mg/kg in a manner consistent with the condition of thepatient. Preferably the oral dose would be about 0.5 to about 20 mg/kg.

No unacceptable toxicological effects are expected when compounds of thepresent invention are administered in accordance with the presentinvention.

Biological Assays

The compounds of this invention may be tested in one of severalbiological assays to determine the concentration of compound which isrequired to have a given pharmacological effect.

Determination of Cathepsin K Proteolytic Catalytic Activity

All assays for cathepsin K were carried out with human recombinantenzyme. Standard assay conditions for the determination of kineticconstants used a fluorogenic peptide substrate, typicallyCbz-Phe-Arg-AMC, and were determined in 100 mM Na acetate at pH 5.5containing 20 mM cysteine and 5 mM EDTA. Stock substrate solutions wereprepared at concentrations of 10 or 20 mM in DMSO with 20 uM finalsubstrate concentration in the assays. All assays contained 10% DMSO.Independent experiments found that this level of DMSO had no effect onenzyme activity or kinetic constants. All assays were conducted atambient temperature. Product fluorescence (excitation at 360 nM;emission at 460 nM) was monitored with a Perceptive Biosystems CytofluorII fluorescent plate reader. Product progress curves were generated over20 to 30 minutes following formation of AMC product.

Inhibition Studies

Potential inhibitors were evaluated using the progress curve method.Assays were carried out in the presence of variable concentrations oftest compound. Reactions were initiated by addition of enzyme tobuffered solutions of inhibitor and substrate. Data analysis wasconducted according to one of two procedures depending on the appearanceof the progress curves in the presence of inhibitors. For thosecompounds whose progress curves were linear, apparent inhibitionconstants (K_(i,app)) were calculated according to equation 1 (Brandt etal., Biochemitsry, 1989, 28, 140):

v=V _(m) A/[K _(a)(1+I/K _(i, app))+A]  (1)

where v is the velocity of the reaction with maximal velocity V_(m), Ais the concentration of substrate with Michaelis constant of K_(a), andI is the concentration of inhibitor.

For those compounds whose progress curves showed downward curvaturecharacteristic of time-dependent inhibition, the data from individualsets was analyzed to give k_(obs) according to equation 2:

[AMC]=v _(ss) t+(v ₀ −v _(ss))[1−exp(−k _(obs) t)]/k _(obs)  (2)

where [AMC] is the concentration of product formed over time i, v₀ isthe initial reaction velocity and v_(ss) is the final steady state rate.Values for k_(obs) were then analyzed as a linear function of inhibitorconcentration to generate an apparent second order rate constant(k_(obs)/inhibitor concentration or k_(obs)/[I]) describing thetime-dependent inhibition. A complete discussion of this kinetictreatment has been fully described (Morrison et al., Adv. Enzymol.Relar. Areas Mol. Biol., 1988, 61, 201).

Human Osteoclast Resorption Assay

Aliquots of osteoclastoma-derived cell suspensions were removed fromliquid nitrogen storage, warmed rapidly at 37° C. and washed ×1 inRPMI-1640 medium by centrifugation (1000 rpm, 5 min at 4° C.). Themedium was aspirated and replaced with murine anti-HLA-DR antibody,diluted 1:3 in RPMI-1640 medium, and incubated for 30 min on ice Thecell suspension was mixed frequently.

The cells were washed ×2 with cold RPMI-1640 by centrifugation (1000rpm, 5 min at 4° C.) and then transferred to a sterile 15 mL centrifugetube. The number of mononuclear cells were enumerated in an improvedNeubauer counting chamber.

Sufficient magnetic beads (5/mononuclear cell), coated with goatanti-mouse IgG, were removed from their stock bottle and placed into 5mL of fresh medium (this washes away the toxic azide preservative). Themedium was removed by immobilizing the beads on a magnet and is replacedwith fresh medium.

The beads were mixed with the cells and the suspension was incubated for30 min on ice. The suspension was mixed frequently. The bead-coatedcells were immobilized on a magnet and the remaining cells(osteoclast-rich fraction) were decanted into a sterile 50 mL centrifugetube. Fresh medium was added to the bead-coated cells to dislodge anytrapped osteoclasts. This wash process was repeated ×10. The bead-coatedcells were discarded.

The osteoclasts were enumerated in a counting chamber, using alarge-bore disposable plastic pasteur pipette to charge the chamber withthe sample. The cells were pelleted by centrifugation and the density ofosteoclasts adjusted to 1.5×10⁴/mL in EMEM medium, supplemented with 10%fetal calf serum and 1.7 g/liter of sodium bicarbonate. 3 mL aliquots ofthe cell suspension (per treatment) were decanted into 15 mL centrifugetubes. These cells were pelleted by centrifugation. To each tube 3 mL ofthe appropriate treatment was added (diluted to 50 uM in the EMEMmedium). Also included were appropriate vehicle controls, a positivecontrol (87MEM1 diluted to 100 ug/mL) and an isotype control (IgG2adiluted to 100 ug/mL). The tubes were incubate at 37° C. for 30 min.

0.5 mL aliquots of the cells were seeded onto sterile dentine slices ina 48-well plate and incubated at 37° C. for 2 h. Each treatment wasscreened in quadruplicate. The slices were washed in six changes of warmPBS (10 mL/well in a 6-well plate) and then placed into fresh treatmentor control and incubated at 37° C. for 48 h. The slices were then washedin phosphate buffered saline and fixed in 2% glutaraldehyde (in 0.2Msodium cacodylate) for 5 min., following which they were washed in waterand incubated in buffer for 5 min at 37° C. The slices were then washedin cold water and incubated in cold acetate buffer/fast red garnet for 5min at 4° C. Excess buffer was aspirated, and the slices were air driedfollowing a wash in water.

The TRAP positive osteoclasts were enumerated by bright-field microscopyand were then removed from the surface of the dentine by sonication. Pitvolumes were determined using the Nikon/Lasertec ILM21W confocalmicroscope.

General

Nuclear magnetic resonance spectra were recorded at either 250 or 400MHz using, respectively, a Bruker AM 250 or Bruker AC 400 spectrometer.CDCl₃ is deuteriochioroform, DMSO-d₆ is hexadeuteriodimethylsulfoxide,and CD₃OD is tetradeuteriomethanol. Chemical shifts are reported inparts per million (d) downfield from the internal standardtetramethylsilane. Abbreviations for NMR data are as follows: s=singlet,d=doublet, t=triplet, q=quartet, m=multiplet, dd=doublet of doublets,dt=doublet of triplets, app=apparent, br=broad. J indicates the NMRcoupling constant measured in Hertz. Continuous wave infrared (IR)spectra were recorded on a Perkin-Elmer 683 infrared spectrometer, andFourier transform infrared (FTIR) spectra were recorded on a NicoletImpact 400 D infrared spectrometer. IR and FTIR spectra were recorded intransmission mode, and band positions are reported in inversewavenumbers (cm⁻¹). Mass spectra were taken on either VG 70 FE, PE SyxAPI m, or VG ZAB HF instruments, using fast atom bombardment (FAB) orelectrospray (ES) ionization techniques. Elemental analyses wereobtained using a Perkin-Elmer 240C elemental analyzer. Melting pointswere taken on a Thomas-Hoover melting point apparatus and areuncorrected. All temperatures are reported in degrees Celsius.

Analtech Silica Gel GF and E. Merck Silica Gel 60 F-254 thin layerplates were used for thin layer chromatography. Both flash and gravitychromatography were carried out on E. Merck Kieselgel 60 (230-400 mesh)silica gel.

Where indicated, certain of the materials were purchased from theAldrich Chemical Co., Milwaukee, Wis., Chemical Dynamics Corp., SouthPlainfield, N.J., and Advanced Chemtech, Louisville, Ky.

EXAMPLES

In the following synthetic examples, temperature is in degreesCentigrade (° C.). Unless otherwise indicated, all of the startingmaterials were obtained from commercial sources. Without furtherelaboration, it is believed that one skilled in the art can, using thepreceding description, utilize the present invention to its fullestextent. These Examples are given to illustrate the invention, not tolimit its scope. Reference is made to the claims for what is reserved tothe inventors hereunder.

Example 1

Preparation of(2S,1′S)-2-(benzyloxycarbonyl)amino-N-1′-(2-carboethoxythiazol-4-yl)-3′-methylbutyl]methylpentanamide

a) N-benzyloxycarbonyl-L-leucinyl-L-leucinyl bromomethylketone

1-Methyl-3-nitro-1-nitrosoguanidine (5.9 g, 40.11 mmol) in ether (200mL) is cooled to 0° C. 40% potassium hydroxide is added slowly and thediazomethane is allowed to collect in the ether solution for 30 minutesat 0° C.

N-Cbz-L-Leucinyl-L-Leucine (Bachem) (4.0 g, 10.58 mmol) is stirred intetrahydrofuran at −40° C. N-methylmorpholine (1.07 g, 10.58 mmol, 1.16mL) and isobutyl chloroformate (1.45 g, 10.58 mmol, 1.38 mL) are added.The mixture is stirred at −40° C. for 15 minutes and then filtered intoa cold flask to remove precipitated salts. To the filtered solution isadded an excess of the previously prepared diazomethane solution and themixture is allowed to stand at 0° C. for 16 h. An excess of 30% HBr inacetic acid is added at 0° C. and the solution is then washedsuccessively with 1.0N citric acid, saturated aqueous sodium bicarbonate(carefully), and brine. The solution is dried over sodium sulfate,filtered, and evaporated to give the title compound as a white solid(4.10 g). ¹H NMR (400 MHz, CDCl₃) δ 7.34 (m, 5H), 6.51 (d, 1H), 5.15 (d,1H), 5.10 (s, 2H), 4.78 (m, 1H), 4.20 (m, 1H), 4.04 (dd, 2H), 1.63 (m,6H), 0.93 (m, 12H).

b)(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-carboethoxythiazol-4-yl)-3′-methylbutyl]-4-methylpentanamide

The compound of Example 1(a) (2.0 g, 4.4 mmol) and ethyl thiooxamate(0.59 g, 4.4 mmol) were refluxed in ethanol for 4 h. The solvent wasevaporated and the residue chromatographed (silica gel, 2.5%methanol/dichloromethane) to give the title compound as a white solid(1.46 g). ¹H NMR (400 MHz, CDCl₃) δ 7.32 (s, 1H), 7.21 (m, 5H), 6.40 (d,1H), 5.13 (dd, 1H), 5.02 (s, 2H), 4.41 (q, 2H), 4.06 (m, 1H), 1.71 (m,2H), 1.47 (m, 4H), 1.33 (t, 3H), 0.73 (m, 12H).

Example 2

Preparation of(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-carboxythiazol-4-yl-3′-methylbutyl]-4-methylpentanamide

The compound of Example 1(c) (0.92 g, 1.88 mmol) was stirred intetrahydrofuran at 0° C. with 1.0N sodium hydroxide. After stirring for1 h, the solution was quenched with 1.0N citric acid and extracted threetimes with dichloromethane. The combined organic extracts wereevaporated in vacuo to give the title compound as a white solid (0.844g). ¹H NMR (400 MHz, CDCl₃) δ 7.40 (s, 1H), 7.23 (m, 5H), 6.89 (d, 1H),5.22 (d, 1H), 5.14 (dd, 1H), 5.02 (s, 2H), 4.15 (m, 1H), 1.67 (m, 2H),1.44 (m, 4H), 0.81 (m, 12H).

Example 3

Preparation of(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-carboxamidothiazol-4-yl)-3′-methylbutyl]-4-methylpentanamide

The compound of Example 2 (0.408 g, 0.88 mmol) in tetrahydrofuran wascooled to −40° C. and treated with N-methylmorpholine (0.185 g, 1.85mmol, 0.2 mL) and isobutyl chloroformate (0.12 g, 0.88 mmol, 0.11 mL).The mixture was stirred at −40° C. for 15 minutes and then ammonia wasbubbled through the solution for several minutes. The mixture wasallowed to warm to room temperature and was then diluted with ethylacetate and washed successively with 1.0N citric acid, 5% aqueous sodiumbicarbonate, and brine. The organic solution was dried over magnesiumsulfate, filtered, and evaporated to a residue which was chromatographed(silica gel, 3% methanol/dichloromethane) to give the title compound asa white solid (0.245 g). ¹H NMR (400 MHz, CDCl₃) δ 7.22 (m, 5H), 7.04(s, 1H), 6.40 (br s, 1H), 5.51 (br s, 1H), 5.09 (m, 1H), 5.02 (dd, 2H),4.07 (m, 1H), 1.66-1.42 (m, 6H), 0.82 (m, 12H).

Example 4

Preparation of(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-cyanothiazol-4-yl)-3′-methylbutyl]-4-methylpentanamide

The compound of Example 3 (0.185 g, 0.4 mmol) was dissolved indichloromethane, cooled to 0° C. and treated with TFAA (0.093 g, 0.44mmol, 0.06 mL) and pyridine (0.07 g, 0.88 mmol, 0.07 mL). After 3 h, themixture was poured into a solution of saturated aqueous sodiumbicarbonate and extracted with dichloromethane. The organic extractswere washed with 5% hydrochloric acid and brine, dried over magnesiumsulfate, filtered, and evaporated to an oil which was chromatographed(silica gel, 40% ethyl acetate/hexane) to give the title compound as awhite solid (0.095 g). ¹H NMR (400 MHz, CDCl₃) δ 7.44 (s, 1H), 7.29 (s,5H), 6.51 (brd, 1H), 5.14 (m, 1H), 5.07 (s, 2H), 4.11 (m, 1H), 1.78-1.41(m, 6H), 0.83 (m, 12H).

Example 5

Preparation of(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-[2-(N′-benzylcarboxamido)thiazol-4-yl]-3′-methylbutyl]-4-methylpentanamide

To a solution of the compound of Example 2 (0.12 g, 0.26 mmol) indichloromethane under argon at room temperature is added benzylamine(0.03 g, 0.29 mmol, 0.03 mL), BOP reagent (0.115 g, 0.26 mmol), andtriethyl amine (0.026 g, 0.26 mmol, 0.04 mL) which is allowed to stirfor 16 h. The solution is washed with water, then brine and the organiclayer is dried over magnesium sulfate, filtered, and evaporated to givea residue which was chromatographed (silica gel, 40% ethylacetate/hexane) to give the title compound as a white solid (0.065 g).¹H NMR (400 MHz, CDCl₃) δ 7.56 (br s, 1H), 7.33 (m, 10H), 6.48 (br d,1H), 5.15 (dd, 1H), 5.03 (s, 2H), 4.63 (d, 2H), 4.12 (m, 1H), 1.72-1.40(m, 6H), 0.85 (m, 12H).

Example 6

Preparation of(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-[2-[N′-(3-methylpropyl)carboxamido]thiazol-4-yl]-3′-methylbutyl]-4-methylpentanamide

Following the procedure of Example 5, except substituting isobutylaminefor benzylamine, the title compound was prepared (0.074 g). ¹H NMR (400MHz, CDCl₃) a 7.27 (s, 5H), 7.19 (s, 1H), 6.38 (br d, 1H), 5.09 (m, 1H),5.01 (s, 2H), 4.07 (m, 1H), 3.20 (dd, 2H), 1.83 (m, 1H), 1.69-1.40 (m,6H), 0.90 (d, 6H), 0.81 (m, 12H).

Example 7

Preparation of(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-[2-[N′-(2-phenylethyl)carboxamido]thiazol-4-yl]-3′-methylbutyl]methylpentanamide

Following the procedure of Example 5, except substituting2-phenylethylamine for benzylamine, the title compound was prepared(0.070 g). ¹H NMR (400 MHz, CDCl₃) δ 7.30-7. 11 (m, 11H), 6.35 (br d,11H), 5.09 (m, 1H), 5.01 (s, 2H), 4.05 (m, 1H), 3.64 (m, 2H), 2.87 (t,2H), 1.69-1.40 (m, 6H), 0.80 (m, 12H).

Example 8

Preparation of(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(4-carboethoxythiazol-2-yl)-3′-methylbutyl]-4-methylpentanamide

a) N-tert-butoxycarbonyl-(L)-leucinamide

To a solution of N-tert-butoxycarbonyl-(L)-leucine (Advanced Chemtech)(5.0 g, 20.0 mmol ) in dry THF (100 mL) at −40° C. was added isobutylchloroformate (2.7 g, 20.0 mmol) and N-methylmorphiline (4.2 g, 42mmol). After 15 minutes of stirring, ammonia was bubbled through themixture for an additional 15 minutes, then warned to room temperatureand allowed to stir for 2 hours. Mixture filtered and filtrateconcentrated in vacuo to yield title compound as a white solid (4.9 g,19.7 mmol). ¹H NMR (400 MHz, CDCl₃) δ 6.38 (br s, 1H ), 5.79 (br s, 1H),5.04 (br d, 1H), 4.13 (m, 1H), 1.71-1.49 (m, 3H), 1.39 (s, 9H), 0.92(dd, 6H).

b) N-tert-butoxycarbonyl-L-leucinethioamide

To a stirring solution of the compound of Example 8(a) (2.38 g, 10.35mmol) in dry THF was added Lawessons reagent (2.51 g, 6.21 mmol) and themixture was stirred at room temperature under argon overnight. Thesolvent was evaporated and the residue chromatographed (silica gel, 2.5%methanol/dichloromethane) to give the title compound as a white solid(2.3 g). ¹H NMR (400 MHz, CDCl₃) δ 8.54 (br s, 1H), 7.97 (br s, 1H),5.28 (br d, 1H), 4.52 (m, 1H), 1.72-1.58 (m, 3H), 1.40 (s, 9H), 0.92 (d,6H).

c)(1S)-1-(tert-butoxycarbonyl)amino-1-(4-carboethoxythiazol-2-yl)-3-methylbutane

The compound of Example 8(b) (2.40 g, 9.76 mmol) was stirred in dryacetone (20 mL) under argon at −10° C. Ethylbromopyruvate (2.12 g, 10.73mmol, 1.35 mL) was added and stirred for 1 h at −10° C. The solution waspoured into a well stirred mixture of chloroform and water and thensaturated with sodium bicarbonate. The organic phase was separated andthe aqueous layer extracted with chloroform. The combined organicextracts were dried over MgSO₄, filtered, and evaporated to an oil. Theoily residue was treated with TFAA (2.19 g, 10.73 mmol, 1.5 mL) andpyridine (1.70 g, 21.47 mmol, 1.75 mL) in dichloromethane for 1 h at−20° C. Excess solvent was removed in vacuo and the residue wasdissolved in dichloromethane. The solution was washed with saturatedaqueous sodium bicarbonate and 1.0N KHSO₄ until pH 7. The solution wasdried over sodium sulfate, filtered, and evaporated to an oil which waschromatographed (4% methanol/dichloromethane) to give the title compoundas a tan solid (1.2 g). ¹H NMR (400 MHz, CDCl₃) δ 7.98 (s, 1H), 5.04 (brd, 1H), 4.95 (m, 1H), 4.31 (q, 2H), 1.88 (m, 1H), 1.63 (m, 2H), 1.40 (s,9H), 1.32 (t, 3H), 0.85 (dd, 6H).

d)(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(4-carboethoxythiazol-2-yl)-3′-methylbutyl]-4-methylpentanamide

The compound of Example 8(c) (1.0 g, 2.92 mmol) was dissolved in neatTFA (1.0 mL) and stirred for 15 minutes. The solution was diluted withmethanol and evaporated in vacuo. A portion of the residue obtained(0.36 g, 1.49 mmol) was dissolved in dichloromethane withN-Cbz-L-leucine (0.394 g, 1.49 mmol), BOP reagent (0.66 g, 1.49 mmol),and triethylamine (0.73 g, 7.2 mmol, 1.0 mL) and stirred at roomtemperature for 16 h. The solution was washed with water, then brine anddried over magnesium sulfate, filtered, and evaporated to a residuewhich was chromatographed (silica gel, 40% ethyl acetate/hexane) to givethe title compound as a white solid (0.396 g). ¹H NMR (400 MHz, CDCl₃) δ7.96 (s, 1H), 7.25 (s, 5H), 6.61 (br d, 1H), 5.30 (m, 1H), 5.09 (br d,1H), 5.01 (s, 2H), 4.33 (q, 2H), 4.10 (m, 1H), 1.90-1.58 (m, 6H), 1.29(t, 3H), 0.81 (dd, 12H).

Example 9

Preparation of(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(4-carboxythiazol-2-yl)-3′-methylbutyl]-4-methylpentanamide

Following the procedure of Example 2, except substituting(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(4-carboethoxythiazol-2-yl)-3′-methylbutyl]-4-methylpentanamidefor(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-carboxythiazol-4-yl)-3′-methylbutyl]4-methylpentanamide,the title compound was prepared (0.301 g). ¹H NMR (400 MHz, CDCl₃) δ8.06 (s, 1H), 7.24 (m, 5H), 7.11 (d, 1H), 5.30 (m, 2H), 5.04 (s, 2H),4.16 (m, 1H), 1.88-1.40 (m, 6H), 0.71 (dd, 12H).

Example 10

Preparation of(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(4-carboethoxythiadiazol-2-yl)-3′-methylbutyl]4-methylpentanamide

a) N-tert-butoxycarbonyl-L-leucine Methyl Ester

To a stirring suspension of L-leucine methyl ester hydrochloride(Aldrich) (6.00 g, 33.0 mmol) and di-tert-butyl dicarbonate (7.21 g,33.0 mmol) in THF (35 mL) was added triethylamine (3.34 g, 33.0 mmol,4.60 mL). The mixture was allowed to stir at room temperature for 3 d.The mixture was diluted with ethyl acetate and washed with 1 N HCl (2times), water, and saturated brine, then dried over magnesium sulfate,filtered and concentrated to give the title compound as a colorless oil(8.02 g, 99%). ¹H NMR (400 MHz, CDCl₃) δ 4.88 (d, 1H), 4.33-4.31 (m,1H), 7.73 (s, 3H), 1.75-1.48 (m, 3H), 1.44 (s, 9H), 0.96 (d, 3H), 0.93(d, 3H).

b) N-tert-butoxycarbonyl-L-leucine Hydrazide

To a stirring solution of the compound of Example 10(a) (8.02 g, 32.7mmol) in methanol (250 mL) was added hydrazine hydrate (16.38 g, 327mmol, 15.9 mL). After stirring for 22 h at room temperature, thesolution was concentrated and the residue was azeotroped with toluene toprovide the title compound as a white foam (8.02 g, 100%). ¹H NMR (400MHz, CDCl₃) δ 7.71 (br s, 1H), 4.99 (d, 2H), 4.124.10 (m, 1H), 3.94 (brs, 2H), 1.68-1.49 (m, 3H), 1.44 (s, 9H), 0.95 (d, 3H), 0.92 (d, 3H).

c)(2S)-N-[2-(benzyloxycarbonyl)amino-4-methylpentanoyl]-N′-carboethoxycarbonylhydrazide

To a stirring solution of the compound of Example 10(b) (8.02 g, 32.7mmol) and pyridine (2.85 g, 36.0 mmol, 2.91 mL) in dichloromethane (200mL) was added ethyl oxalyl chloride (4.91 g, 36.0 mmol, 4.02 mL). Afterstirring at room temperature for 2 h, thye solution was washed with 1 NHCl, water, saturated aqueous sodium bicarbonate and saturated brine,then dried over magnesium sulfate, filtered, and concentrated to affordthe title compound as a white foam (9.84 g, 87%). ¹H NMR (400 MHz,CDCl₃) δ 9.32 (br s, 2H), 5.04 (d, 2H), 4.38 (q, 2H), 4.28 (m, 1H),1.77-1.56 (m, 3H), 1.44 (s, 9H), 1.39 (t, 3H), 0.96 (d, 3H), 0.94 (d,3H).

d)(1S)-1-(tert-butoxycarbonyl)amino-1-(4-carboethoxythiadiazol-2-yl)-3-methylbutane

To a stirring solution of the compound of Example 10(c) (2.50 g, 7.24mmol) in toluene (70 mL) was added Lawesson's reagent (1.46 g, 3.62mmol). The mixture was heated at reflux for 3 h. The solution wasdiluted with ether, washed with saturated aqueous sodium bicarbonate andsaturated brine, then dried over magnesium sulfate, filtered andconcentrated to leave a pale yellow oil. The crude material was purifiedby flash chromatography on 75 g of 230-400 mesh silica gel, eluting with1:4 ethyl acetate/hexanes, to provide the tide compound as a pale yellowsolid (1.75 g, 70%). ¹H NMR (400 MHz, CDCl₃) δ 5.19 (m, 1H), 5.13 (d,1H), 4.51 (q, 2H), 1.95 (m, 1H), 1.83-1.73 (m, 2H), 1.44 (s, 9H), 1.00(d, 3H), 0.98 (d, 3H).

e) (1S)-1-amino-1-(4-carboethoxythiadiazol-2-yl)-3-methylbutanebis-trifluoroacetate Salt

To a stirring solution of the compound of Example 10(d) (1.75 g, 5.1mmol) in dichloromethane (40 mL) was added TFA (10 mL). After stirringfor 5 min at room temperature, the solution was concentrated to give thetitle compound as an oily pale yellow solid (2.40 g, 100%). ¹H NMR (400MHz, CDCl₃) δ 9.83 (br s, 4H), 5.20 (m, 1H), 4.51 (q, 2H), 2.07 (m, 2H),1.70 (m, 1H), 1.44 (t, 3H), 1.00 (t, 3H).

f)(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(4-carboethoxythiadiazol-2-yl)-3′-methylbutyl]4-methylpentanamide

To a stirring solution of the compound of Example 10(e) (566.1 mg, 1.20mmol), N-Cbz-L-leucine (250.5 mg, 1.32 mmol),1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (253.3 mg,1.32 mmol) and 1-hydroxybenzotriazole (32.5 mg, 0.24 mmol) in 2.5 mL ofDMF was added triethylamine (243.1 mg, 2.40 mmol, 0.335 mL). Afterstirring at room temperature for 3 d, the mixture was diluted with ethylacetate and washed with water, saturated aqueous sodium bicarbonate andsaturated brine, then dried over magnesium sulfate, filtered andconcentrated to give a yellow oil. The cride material was purified byflash chromatography on 20 g of 230-400 mesh silica gel, eluting with1:2 ethyl acetate/hexanes, to provide the title compound as a whitesolid (271 mg, 46%). ¹H NMR (400 MHz, CDCl₃) δ 7.35 (s, 5H), 6.77 (d,1H), 5.49 (m, 1H), 5.12 (dd, 2H), 4.51 (q, 2H), 4.20 (m, 1H), 1.97 (m,1H), 1.88 (m, 1H), 1.66 (m, 3H), 1.52 (m, 1H), 1.45 (t, 3H), 0.97-0.92(m, 12H).

Example 11

Preparation of(2S,1′S)-2-(benzyloxycarbonyl)amino--N-[1′-(2-carbo-2,2,2-trifluoroethoxythiazol-4-yl)-3′-methylbutyl]-4-methylpentanamide

The compound of Example 2 (0.200 g, 0.433 mmol), 1,1,1 trifluoroethanol(0.052 g, 0.52 mmol, 0.04 mL), pyridine (0.1 ml), and di-t-butyldicarbonate (0.104 g, 0.477 mmol) were stirred in ethyl acetate at roomtemperature for 16 h. The solution was diluted with ethyl acetate andwashed successively with 5% hydrochloric acid, 10% aqueous sodiumbicarbonate, and brine. The organic layer was dried over magnesiumsulfate, filtered, and evaporated to give a residue which waschromatographed (silica gel, 20% ethyl acetate/hexane) to give the titlecompound as a white solid (0.098 g). ¹H NMR (400 MHz, CDCl₃) δ 7.50 (s,1H), 7.36 (s, 5H), 6.64 (d, 1H), 5.22 (m, 2H), 5.09 (s, 2H), 4.73 (m,2H), 4.16 (m, 1H), 1.66-1.41 (m, 6H), 0.87 (m, 12H).

Example 12

Preparation of(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(4-carboethoxyoxadiazol-2-yl)-3′-methylbutyl]-4-methylpentanamide

a)(1S)-1-(tert-butoxycarbonyl)amino-1-(4-carboethoxyoxadiazol-2-yl)-3-methylbutane

To a stirring solution of the compound of Example 10(c) (2.50 g, 7.24mmol) and pyridine (1.49 g, 18.8 mmol, 1.52 mL) in ether (15 mL) wasadded thionyl chloride (1.12 g, 9.41 mmol, 0.69 mL). After stirring atroom temperature for 2 h, the solid was removed by filtration and thefiltrate was concentrated. The residue was dissolved in toluene andheated at reflux. After 12 h, the solution was concentrated to leave abrown oil. The residue was purified by flash chromatography on 175 g of230-400 mesh silica gel, eluting with 1:4 ethyl acetate/hexanes, to givethe title compund as a pale yellow oil (0.84 g, 35%). ¹H NMR (400 MHz,CDCl₃) δ 5.14 (m, 1H), 5.03 (br d, 1H), 4.52 (q, 2H), 1.78-1.70 (m, 3H),1.44 (s, 9H), 0.99 (d, 6H).

b) (1S)-1-amino-1-(4-carboethoxyoxadiazol-2-yl)-3-methylbutane

Following the procedure of Example 10(e), except substituting(1S)-1-(tert-butoxycarbonyl)amino-1-(4-carboethoxyoxadiazol-2-yl)-3-methylbutanefor(1S)-1-(tert-butoxycarbonyl)amino-1-(4-carboethoxythiadiazol-2-yl)-3-methylbutane,the title compound was prepared (582 mg, 100%). ¹H NMR (400 MHz, CDCl₃)δ 4.99 (t, 1H), 4.52 (q, 2H), 2.10-2.02 (m, 2H), 1.77-1.70 (m, 1H), 1.44(t, 3H), 1.00 (t, 6H).

c)(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(4-carboethoxyoxadiazol-2-yl)-3′-methylbutyl]4methylpentanamide

Following the procedure of Example 10(f), except substituting(1S)-1-amino-[1-(4-carboethoxyoxadiazol-2-yl)-3-methylbutane for(1S)-1-amino-1-(4-carboethoxythiadiazol-2-yl)-3-methylbutane, the titlecompound was prepared (235 mg, 39%). ¹H NMR (400 MHz, CDCl₃) δ 7.26 (s,5H), 6.64 (s, 1H), 5.45-5.39 (m, 1H), 5.12 (m, 3H), 4.52 (q, 2H), 4.20(m, 1H), 1.81 (m, 2H), 1.68-1.64 (m, 3H), 1.54-1.50 (m, 1H), 1.46 (t,3H), 0.97-0.92 (m, 12H).

Example 13

Preparation of(2S,1′S)-2-(benzyloxycarbonyl-L-leucinyl)amino-N-[1′-(4-carboethoxythiazol-2-yl)-3′-methylbutyl]-4-methylpentanamide

The compound of Example 8(c) (160.7 mg, 0.47 mmol) was dissolved in neatTFA (1.0 mL) and stirred for 15 minutes. The solution was diluted withmethanol and evaporated to dryness. The residue was dissolved in DMF (2mL) and to the resulting solution was added N-Cbz-L-leucinyl-L-leucine(194.0 mg, 0.52 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride (99.0 mg, 0.52 mmol) and 1-hydroxybenzotriazole (13.0 mg,0.094 mmol) and triethylamine (94.7 mg, 0.936 mmol, 0.13 mL). Afterstirring at room temperature for 24 h, the mixture was diluted withethyl acetate and washed with water, saturated aqueous sodiumbicarbonate and saturated brine, then dried over magnesium sulfate,filtered and concentrated. The residue was purified by flashchromatography on 230-400 mesh silica gel, eluting with ethylacetate/hexanes, to provide the title compound (0.146 g). ¹HNMR(400 MHz,CDCl₃) δ 8.04 (s, 1H), 7.33 (m, 5H), 7.14 (d, 1H), 6.61 (d, 1H), 5.37(m, 2H), 5.08 (m, 2H), 4.47 (m, 1H), 4.39 (q, 2H), 4.18 (m, 1H),1.98-1.45 (m, 9H), 1.38 (t, 3H), 0.94-0.86 (m, 18H).

Example 14

Preparation of(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1-(4-carboxamidooxadiazol-2-yl)-3′-methylbutyl]-4-methylpentanamide

Ammonia was bubbled through a solution of the compound of Example 12(96.8 mg, 0.2 mmol) in ethanol (2 mL) for 5 inm. After stirring anadditional 5 min, the solution was concentrated to give the titlecompound as a white solid (91.2 mg, 98%). ¹H NMR (400 MHz, CDCl₃/CD₃OD)δ 7.29 (s, 5H), 5.90 (d, 1H), 5.30 (t, 1H), 5.04 (s, 2H), 4.15 (m, 1H),1.76 (m, 2H), 1.59-1.43 (m, 4H), 0.92-0.85 (m, 12H).

Example 15

Preparation of(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-carboethoxythiazol-4-yl)-3′-methylbutyl]-3-phenylpropanamide

a) N-(9-fluorenylmethoxycarbonyl)-L-leucinyl Bromomethyl Ketone

Following the procedure of Example 1(a), except substitutingN-(9-fluorenylmethoxycarbonyl)-L-leucine forN-benzyloxycarbonyl-L-leucinyl-L-leucine, the title compound wasprepared (5.6 g). ¹H NMR (400 MHz, CDCl₃) δ 7.71 (d, 2H), 7.51 (d, 2H),7.34 (dd, 2H), 7.22 (dd, 2H), 5.08 (d, 1H), 4.53 (m, 1H), 4.36 (dd, 2H),4.13 (dd, 2H), 3.89 (dd, 2H), 1.62-1.41 (m, 3H), 0.88 (m, 6H).

b)(1S)-1-(2-carboethoxythiazol-4yl)-1-(9-fluorenylmethoxycarbonyl)amino-3-methylbutane

Following the procedure of Example 1(b), except substitutingN-(9-fluorenylmethoxycarbonyl)-L-leucinyl bromomethyl ketone forN-Benzyloxycarbonyl-L-leucinyl-L-leucinyl bromomethylketone, the titlecompound was prepared (4.13 g). ¹H NMR (400 MHz, CDCl₃) δ 7.72 (d, 2H),7.49 (d, 2H), 7.32 (dd, 2H), 7.22 (dd, 2H), 7.19 (s, 1H), 5.31 (d, 1H),4.88 (m, 1H), 4.40 (q, 2H), 4.28 (d, 2H), 4.08 (t, 1H), 1.62-1.41 (m,3H), 1.36 (t, 3H), 0.88 (m, 6H).

c) (1S)-1-amino-1-(2-carboethoxythiazol-4-yl)-3-methylbutane

The compound of Example 15(b) (0.5 g., 1.1 mmol) was stirred in a 5%piperidine/DMF solution for 10 minutes at room temperature. The solventswere evaporated and the solid obtained was dried in vacuo to give thetitle compound (0.27 g).

d) (2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2carboethoxythiazol4yl)-3′-methylbutyl]-3-phenylpropanamide Following the procedure ofExample 5, except substituting(1S)-1-amino-1-(2-carboethoxythiazol-4-yl)-3-methylbutane forbenzylamine, and N-Cbz-L-phenylalanine for(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-carboxythiazol-4-yl)-3′-methylbutyl]4-methylpentanamide,the title compound was prepared (0.162 g). ¹H NMR (400 MHz, CDCl₃) δ7.27 (m, 5H), 7.11 (s, 1H), 7.04 (m, 5H), 6.12 (d, 1H), 5.24 (d, 1H),5.10 (q, 1H), 5.01 (s, 2H), 4.37 (q, 2H), 4.21 (m, 1H), 2.91 (m, 2H),1.62 (m, 3H), 1.37 (t, 3H), 0.81 (m, 6H).

Example 16

Preparation of(2S,1′S)-2-(benzyloxycarbonyl-L-leucinyl)amino-N-[1′-(2-carboethoxythiazol-4-yl)-3′-methylbutyl]-4-methylpentanamide

Following the procedure of Example 5, except substituting(1S)-1-amino-1-(2-carboethoxythiazol-4-yl)-3-methylbutane forbenzylamine, and N-Cbz-L-leucinyl-L-leucine for(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-carboxythiazol-4-yl)-3′-methylbutyl]4-methylpentanamide,the title compound was prepared (0.098 g.). ¹H NMR (400 MHz, CDCl₃) δ7.39 (s, 1H),-7.25 (m, 5H), 6.87 (d, 1H), 6.49 (d, 1H), 5.30 (d, 1H),5.16 (q, 1H), 4.99 (s, 2H), 4.36 (q, 2H), 4.31 (m, 1H), 4.09 (m, 1H),1.74-1.38 (m, 9H), 1.32 (t, 3H), 0.80 (m, 15H).

Example 17

Preparation of(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(5-mercapto-1,2,4-oxadiazol-3-yl)-3′-methylbutyl]-4-methylpentanamide

a) N-benzyloxycarbonyl-L-leucinyl-L-leucine Methyl Ester

N-Cbz-L-leucine (Chemical Dynamics) (1.32 g, 4.97 mmol), L-leucinemethyl ester hydrochloride (Aldrich) (0.99 g, 5.47 mmol),1-hydroxybenzotriazole (0.14 g, 1.0 mmol) and1-(3-dimethylaminopropyl)-3ethylcarbodiimide hydrochloride (1.05 g, 5.47mmol) were combined, dissolved in 25 nL of DMF and stirred at roomtemperature for 15 h. The solution was diluted with ethyl acetate (250mL) and washed successively with water, 0.1 N HCl, saturated aqeousNaHCO₃ and saturated brine, then dried (MgSO₄), filtered, andconcentrated. The residue was purified by flash chromatography on230-400 mesh silica gel, eluting with 1:3 ethyl acetate hexanes, to givethe title compound as a white solid (1.28 g, 66%). ¹H NMR (400 MHz,CDCl₃) δ 7.37-7.32 (m, 5H), 6.28 (d, 1H), 5.28 (m, 3H), 4.61-4.58 (m,1H), 4.20 (m, 1H), 3.74 (s, 3H), 1.69-1.54 (m, 6H), 0.96-0.92 (m, 12H).

b) N-benzyloxycarbonyl-L-leucinyl-L-leucinylhydrazide

To as stirring solution of the compound of Example 17(a) (1.28 g, 3.26mmol) in 25 mL of methanol was added hydrazine hydrate (1.63 g, 32.6mmol, 1.58 mL) and the solution was allowed to stir at room temperaturefor 15 h. The solution was evaporated to dryness to give the titlecompound as a white solid (1.28 g, 100%). ¹H NMR (400 MHz, CDCl₃) δ 8.05(br s, 1H), 7.35-7.32 (m, 5H), 6.67 (d, 1H), 5.50 (d, 1H), 5.11 (s, 2H),4.46 (m, 1H), 4.21 (m, 1H), 3.88 (br s, 2H), 1.64-1.51 (m, 6H),0.92-0.88 (m, 12H).

c)(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(5-mercapto-1,2,4-oxadiazol-3-yl)-3′-methylbutyl]4methylpentanamide

To a stirring solution of the compound of Example 17(b) (0.3 g, 0.76mmol) in 1.5 mL of chloroform was added triethylamine (0.155 g, 1.53mmol, 0.213 mL) and thiophosgene (0.088 g, 0.76 mmol, 0.058 mL). Thesolution was heated at reflux for 3 h, then cooled to room temperature.The mixture was diluted with ethyl acetate, washed with water andsaturated brine, dried (MgSO₄), filtered, and concentrated. The residuewas purified by flash chromatography on 230-400 mesh silica gel, elutingwith 11% methanol in dichloromethane, to give the title compound as awhite solid (0.20 g, 61%). ¹H NMR (400 MHz, CDCl₃) δ 7.36 (m, 6H), 6.85(d, 1H), 5.37 (d, 1H), 5.14 (m, 3H), 4.24 (m, 1H), 1.65 (m, 6H),0.95-0.87 (m, 12H).

Example 18

Preparation of(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-mercaptothiazol-4-yl)-3′-methylbutyl]-4-methylpentanamide

The compound of Example 1(a) (1.0 g, 2.2 mmol) and ammoniumdithiocarbamate (0.25 g., 2.2 mmol) were dissolved in ethanol and heatedto 55° C. for 13 hours. The solvent was evaporated and the residuechromatographed (silica gel, 20% ethyl acetate/hexane) to give the titlecompound as a white solid (0.58 g). ¹H NMR (400 MHz, CDCl₃) δ 7.24 (m,5H), 7.10 (s, 1H), 6.33 (s, 1H), 6.00 (d, 1H), 5.11 (q, 2H), 4.94 (m,1H), 4.05 (m, 1H), 1.49 (m, 6H), 0.78 (m, 12H).

Example 19

Preparation of(2S)-2-(benzyloxycarbonyl)amino-N-(4-carboethoxythiazol-2-yl)methyl-4-methylpentanamide

a) 1-(tert-butoxycarbonyl)amino-1-(4-carboethoxythiazol-2-yl)methane

Following the procedure of Example 8(a)-8(c), except substitutingN-tert-butoxycarbonylglycine for N-tert-butoxycarbonyl-(L)-leucine instep (a), the title compound was prepared (1.9 g, 58% overall). ¹H NMR(400 MHz, CDCl₃) δ 8.11 (s, 1H), 5.31 (s, 1H), 4.56 (d, 2H), 4.43 (q,2H), 1.45 (s, 9H), 1.42 (t, 3H).

b)(2S)-2-(benzyloxycarbonyl)amino-N-(4-carboethoxythiazol-2-yl)methyl-4-methylpentanamide

Following the procedure of Example 13, except substituting1-(tert-butoxycarbonyl)amino-1-(4-carboethoxythiazol-2-yl)methane for(1S)-1-(tert-butoxycarbonyl)amino-1-(4-carboethoxythiazol-2-yl)-3-methylbutane,and N-Cbz-L-leucine for N-Cbz-L-leucinyl-L-leucine, the title compoundwas prepared (0.120 g, 32%). MS (MH⁺): 434.2.

Example 20

Preparation of(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-benzyloxycarbonylthiazol-4-yl)-3′-methylbutyl]-4-methylpentanamide

The compound of Example 2 (0.105 g., 0.22 mmol) was dissolved indichloromethane and treated with1-(3-dimethylaminopropyl)-3-ethylcarbodiimide methiodide (0.062 g., 0.22mmol) and benzyl alcohol (0.03 mL, 0.22 mmol). The mixture was allowedto stir at room temperature for 4 hours and the solvents were evaporatedand the residue obtained was cheromatographed (silica gel, 30% ethylacetate/hexane) to give the title compound as a white solid (0.04 g). ¹HNMR (400 MHz, CDCl₃) δ 7.37 (s, 1H), 7.26 (m, 10H), 6.50 (d, 1H), 5.33(s, 2H), 5.11 (q, 2H), 5.09 (m, 1H), 4.99 (s, 2H), 4.04 (m, 1H), 1.49(m, 6H), 0.78 (m, 12H).

Example 21

Preparation of(2S,1′S)-2-(benzyloxycarbonylamino-4-methyl-N-[3′-methyl-1′-(2-phenoxycarbonylthiazol-4-yl)butyl]pentanamide

Following the procedure of Example 20, except substituting phenol forbenzyl alcohol, the title compound was prepared (0.075 g). ¹H NMR (400MHz, CDCl₃) δ 7.41 (s, 1H), 7.26 (m, 10H), 6.49 (d, 1H), 5.20 (m, 1H),5.04 (m, 1H), 5.00 (s, 2H), 4.08 (m, 1H), 1.49 (m, 6H), 0.82 (m, 12H).

Example 22

Preparation of(2S,1′S)-2-(benzyloxycarbonyl)amino-4-methyl-N-[3‘-methyl-1′-(2-(2-methylpropyloxycarbonyl)thiazol-4-yl)butyl]pentanamide

Following the procedure of Example 20, except substituting isobutylalcohol for benzyl alcohol, the title compound was prepared (0.075 g).¹H NMR (400 MHz, CDCl₃) δ 7.25 (m, 6H), 6.50 (d, 1H), 5.11 (q, 2H), 5.09(m, 1H), 4.99 (s, 2H), 4.11 (d, 2H), 3.91 (m, 1H), 2.02 (m, 1H),1.70-1.39 (m, 6H), 0.82 (d, 6H), 0.78 (m, 12H).

Example 23

Preparation of(2R,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(4-carboethoxythiazol-2-yl)ethyl]-4-methylpentanamide

Following the procedure of Example 19, except substitutingN-tert-butoxycarbonyl-L-alanine for N-tert-butoxycarbonylglycine in step(a), and N-Cbz-D-leucine for N-Cbz-L-leucine in step (b), the titlecompound was prepared as white solid (0.135 g, 36%). MS (MH⁺): 448.2.

Example 24

Preparation of(2R,1′R-2-(benzyloxycarbonyl)amino-N-[1′-(4carboethoxythiazol-2-yl)ethyl]-4-methylpentanamide

Following the procedure of Example 19, except substitutingN-tert-butoxycarbonyl-D-alanine for N-tert-butoxycarbonylglycine in step(a), and N-Cbz-D-leucine for N-Cbz-L-leucine in step (b), the titlecompound was prepared as white solid (0.110 g, 29%). MS (MH⁺): 448.2.

Example 25

Preparation of(2S,1′S)-N-[1′-(2-aminothiazol-4-yl)-3′-methylbutyl]-2-(benzyloxycarbonyl)amino-4-methylpentanamide

To a stirring solution of the compound of Example 1(a) (0.85 g, 1.87mmol) in 4 mL of ethanol was added thiourea (0.142 g, 1.87 mmol). Thesolution was allowed to stir at room temperature for 90 min. Thesolution was concentrated, the residue was dissolved in ethyl acetateand washed with saturated aqeous NaHCO₃ and saturated brine, then dried(MgSO₄), filtered, and concentrated. The residue was purified by flashchromatography on 230-400 mesh silica gel, eluting with 1:1 ethylacetate/hexanes, to give the title compound as a white solid (0.64 g,78%). ¹H NMR (400 MHz, CDCl₃) δ 7.36 (m, 5H), 6.30 (m, 2H), 5.12 (m,3H), 4.95-4.91 (m, 3H), 4.16 (m, 1H), 1.63 (m, 4H), 1.49 (m, 2H),0.93-0.89 (m, 12H).

Example 26

Preparation of(1S)-N-[4-[(1-benzyloxycarbonylamino)-3-methylbutyl]thiazol-2-ylcarbonyl]-N′-(N-benzyloxycarbonyl-L-leucinyl)hydrazide

a) N-benzyloxycarbonyl-L-leucinyl Bromomethyl Ketone

1-methyl-3-nitro-1-nitrosoguanidine (6.65 g, 45.2 mmol) in ether (225mL) is cooled to 0° C. 40% sodium hydroxide is added slowly and thediazomethane is allowed to collect in the ether solution for 30 minutesat 0° C. The ether solution is then decanted and left at 0° C.

N-Cbz-L-leucine (2.10 g, 7.6 mmol) was dissolved in THF (10 mL), cooledto −40° C., and 4-methylmorpholine (0.77 g, 7.6 mmol, 0.83 mL) wasadded, followed by dropwise addition of isobutyl chloroformate (1.04 g,7.6 mmol, 0.98 mL). After 15 min, the solution was filtered into thepreviously prepared 0° C. solution of ethereal diazomethane. Theresulting solution was allowed to stand at 0° C. for 23 h. HBr (30% inacetic acid) (45.2 mmol, 9 mL) was added and the resulting solution wasstirred at 0° C. for 5 min, then washed sequentially with 0.1 N HCl,saturated aqueous NaHCO₃ and saturated brine, then dried (MgSO₄),filtered and concentrated to give the title compound as a colorless oil(2.43 g, 94%).

b)(1S)-1-benzyloxycarbonylamino-1-(2-carboethoxythiazol-4yl)-3-methylbutane

A solution of the compound of Example 26(a) (1.57 g, 4.58 mmol) andethyl thiooxamate (0.61 g, 4.58 mmol) in ethanol (10 mL) was heated atreflux for 4 h. The solution was then cooled, concentrated and theresidue was purified by flash chromatography on 230-400 mesh silica gel,eluting with 1:4 ethyl acetate/hexanes, to give the title compound as ayellow oil (1.0 g, 58%). ¹H NMR (400 MHz, CDCl₃) δ 7.41 (s, 1H),7.34-7.31 (m, 5H), 5.40 (d, 1H), 5.10 (d, 1H), 5.05 (d, 1H), 4.98 (q,1H), 4.48 (q, 2H), 1.80-1.76 (m, 2H), 1.57-1.53 (m, 1H), 1.44 (t, 3H),0.95 (d, 3H), 0.93 (d, 3H).

c)(1S)-1-benzyloxycarbonylamino-1-(2-hydrazinocarbonylthiazol-4-yl)-3-methylbutane

A solution of the compound of Example 26(b) (0.30 g, 0.8 mmol) andhydrazine hydrate (0.40 g, 8.0 mmol, 0.39 mL) in ethanol (8 mL) wasallowed to stir at room temperature for 2 h. The solution was thenconcentrated to yield the title compound as a white foam (0.28 g, 98%).¹H NMR (400 MHz, CDCl₃) δ 8.29 (s, 1H), 7.37-7.35 (m, 5H), 5.18 (d, 1H),5.09 (dd, 2H), 4.95 (q, 1H), 4.07 (d, 2H), 1.71 (t, 2H), 1.55 (m, 1H),0.96 (d, 3H), 0.94 (d, 3H).

d)(1S)-N-[4[(1-benzyloxycarbonylamino)-3-methylbutyl]thiazol-2-ylcarbonyl]-N′-(N-benzyloxycarbonyl-L-leucinyl)hydrazide

A solution of the compound of Example 26(c) (100 mg, 0;28 mmol),N-Cbz-L-leucine (80.5 mg, 0.30 mmol),1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (58.2 mg,0.30 mmol) and 1-hydroxybenzotriazole (7.5 mg, 0.06 mmol) in DMF (0.6mmol) was allowed to stir at room temperature for 18 h. The solution wasdiluted with ethyl acetate and washed successively with water, 0.1 NHCl, saturated aqueous NaHCO₃ and saturated brine, then dried (MgSO₄),filtered and concentrated. The residue was purified by flashchromatography on 230-400 mesh silica gel, eluting with 1:1 ethylacetate/hexanes, to provide the title compound as a white solid (111.4mg, 66%). mp 110-112° C.

Example 27

Preparation ofN-benzyloxycarbonyl-L-leucinyl-N′-benzyloxycarbonyl-L-leucinyl-L-leucinylhydrazide

a) N-benzyloxycarbonyl-L-leucinyl-L-leucine Methyl Ester

Following the procedure of Example 26(d), except L-leucine methyl esterhydrochloride for(1S)-1-benzyloxycarbonylamino-1-(2-hydrazinocarbonylthiazol-4-yl)-3-methylbutane,the title compound was prepared as a white solid (1.28 g, 66%). ¹H NMR(400 MHz, CDCl₃) δ 7.37-7.32 (m, 5H), 6.28 (d, 1H), 5.28 (m, 3H),4.61-4.58 (m, 1H), 4.20 (m, 1H), 3.74 (s, 3H), 1.69-1.54 (m, 6H),0.96-0.92 (m, 12H).

b) N-benzyloxycarbonyl-L-leucinyl-L-leucinylhydrazide

Following the procedure of Example 26(c), except substitutingN-benzyloxycarbonyl-L-leucinyl-L-leucine methyl ester for(1S)-1-benzyloxycarbonylamino-1-(2-carboethoxythiazol-4-yl)-3-methylbutane,the title compound was prepared as a white solid (1.28 g, 100%). ¹H NMR(400 MHz, CDCl₃) δ 8.05 (br s, 1H), 7.35-7.32 (m, 5H), 6.67 (d, 1H),5.50 (d, 1H), 5.11 (s, 2H), 4.46 (m, 1H), 4.21 (m, 1H), 3.88 (br s, 2H),1.64-1.51 (m, 6H), 0.92-0.88 (m, 12H).

c)N-benzyloxycarbonyl-L-leucinyl-N′-benzyloxycarbonyl-L-leucinyl-L-leucinylhydrazide

Following the procedure of Example 26(d), except substitutingN-benzyloxycarbonyl-L-leucinyl-L-leucinylhydrazide for(1S)-1-benzyloxycarbonylamino-1-(2-hydrazinocarbonylthiazol-4yl)-3-methylbutane,the title compound was prepared as a white solid (0.059 g). MS (M+Na⁺):662.1.

Example 28

Preparation of(1S)-N-[2-[(1-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-(N-benzyloxycarbonyl-L-leucinyl)hydrazide

a) N-tert-butoxycarbonyl-(L)-leucinamide

To a solution of N-tert-butoxycarbonyl-(L)-leucine (7.0 g, 28.1 mmol )in dry THF (100 mL) at −40° C. was added isobutylchloroformate (3.8 g,28.1 mmol) and N-methylmorphiline (6.0, 59 mmol). After 15 minutes ofstirring, ammonia was bubbled through the mixture for an additional 15minutes, then warmed to room temperature and allowed to stir for 2hours. Mixture filtered and filtrate concentrated in vacuo to yieldtitle compound as a white solid (6.5, 28.0 mmol). ¹HNMR (400 MHz, CDCl₃)36.38 (br s, 1H), 5.79 (br s, 1H), 5.04 (br d, 1H), 4.13 (m, 1H),1.71-1.49 (m, 3H), 1.39 (s, 9H), 0.92 (dd, 6H).

b) N-tert-butoxycarbonyl-(L)-leucinethioamide

To a stirring solution of the compound of Example 26(a) (6.5, 28.0 mmol)in dry THF was added Lawesson's reagent (6.8 g, 16.9 mmol) and themixture was stirred at room temperature under argon overnight. Thesolvent was evaporated and the residue chromatographed (silica gel, 12%ethyl acetate/hexane) to give the title compound as a white solid (5.4g, 77%). ¹HNMR (400 MHz, CDCl₃) δ 8.54 (br s, 1H), 7.97 (br s, 1H), 5.28(br d, 1H), 4.52 (m, 1H), 1.72-1.58 (m, 3H), 1.40 (s, 9H), 0.92 (m, 6H).

c) (1S)1-(tert-butoxycarbonyl)amino-1-(4-carboethoxythiazol-2-yl)-3-methylbutane

The compound of Example 26(b) (5.4 g, 21.7 mmol) was stirred in dryacetone (100 mL) under argon at −10C. Ethylbromopyruvate (4.7 g, 23.9mmol) was added and stirred for 1 h at −10° C. The solution was pouredinto a well stirred mixture of chloroform and water and then intosaturated sodium bicarbonate solution. The organic phase was separatedand the aqueous layer extracted with chloroform. The combined organicextracts were dried over MgSO₄, filtered and concentrated to an oil. Theoily residue was treated with TFAA (5.0 g, 23.9 mmol) and pyridine (3.8g, 47.8 mmol) in dichloromethane for 1 h at −20° C. Excess solvent wasremoved in vacuo and the residue was dissolved in dichloromethane. Thesolution was washed with saturated aqueous sodium bicarbonate and 1.0NKHSO₄ until pH 7. The solution was dried over magnesium sulfate,filtered and concentrated to an oil which was chromatographed (silicagel, 7.5% ethyl acetate/hexane) to give the title compound as a tansolid (4.5 g, 61%). ¹HNMR (400 MHz, CDCl₃) δ 7.98 (s, 1H), 5.04 (brd,1H), 4.95 (m, 1H), 4.31 (q, 2H), 1.88 (m, 1H), 1.63 (m, 2H), 1.40 (s,9H), 1.32 (t, 3H), 0.85 (dd, 6H).

d)(1S)-1-(Benzyloxycarbonyl)amino-1-(4-carboethoxythiazol-2-yl)-3-methylbutane

The compound of Example 26(c) (0.250 g, 0.731 mmol) was dissolved in TFA(2 mL) and stirred at room temperature for 15 minutes when diluted withmethanol and concentrated in vacuo. The residue was dissolved inmethylene chloride and treated with triethylamine (0.739 g, 7.31 mmol)followed by benzyl chloroformate (1.2 g, 7.311 mmol). The solutionstirred at room temperature for 2 h when partition between ethylacetate/water. The organic layer was washed with brine, collected, dried(MgSO₄) and concentrated to a residue that was chromatographed (silicagel, 15% ethyl acetate/hexane) to give the title compound as an oil(0.198 g, 72%). ¹HNMR (400 MHz, CDCl₃) δ 8.01 (s, 11H), 7.32 (m, 5H),5.51 (br d, 1H), 5.14 (m, 1H), 5.10 (s, 2H), 4.37 (q, 2H), 1.93 (m; 1H),1.81-1.67 (m, 2H), 1.39 (t, 3H), 0.95 (m, 6H)

e)(1S)-N-[2-[(1-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-(N-benzyloxycarbonyl-L-leucinyl)hydrazide

Following the procedure of Example 26(c)-1(d), except substituting(1S)-1-(Benzyloxycarbonyl)amino-1-(4-carboethoxythiazol-2-yl)-3-methylbutanefor(1S)-1-benzyloxycarbonylamino-1-(2-carboethoxythiazolyl)-3-methylbutanein step (c), the title compound was prepared. MS (MH⁺): 610.0.

Example 29

Preparation of2,2′-(N,N′-bis-benzyloxycarbonyl-L-leucinyl)carbohydrazide

To a stirring solution of N-Cbz-L-leucine (Chemical Dynamics Corp.)(2.94 g, 11.1 mmol) in 22 mL of DMF was added carbohydrazide (0.5 g, 5.6mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.13g, 11.1 mmol) and 1-hydroxybenzotriazole (0.3 g, 2.2 mmol). Afterstirring at room temperature for 22 h, the solution was poured into 500mL of water. The precipitate was collected by vacuum filtration andwashed with water (4×150 mL) and dichloromethane (4×150 mL), then driedunder vacuum to provide the title compound as a white solid (1.49 g,46%). MS(ESI): 607.1 (M+Na)⁺.

Example 30

Preparation of 2,2′-(N,N′-bis-cyclohexylacetyl)carbohydrazide

Following the procedure of Example 29, except substitutingcyclohexylacetic acid for N-Cbz-L-leucine, the title compound wasprepared (0.410 g, 73%). MS(ESI): 339.3 (M+H)⁺.

Example 31

Preparation of 2,2′-(N,N′-bis-4-methylpentanoyl)carbohydrazide

Following the procedure of Example 29, except substituting4-methylpentanoic acid for N-Cbz-L-leucine, the title compound wasprepared as a white solid (0.212 g, 44%). MS(ESI): 287.3 (M+H)⁺.

Example 32

Preparation of 2,2′-(N,N′-bis-2-cyclopentylacetyl)carbohydrazide

Following the procedure of Example 29, except substitutingcyclopentylacetic acid for N-Cbz-L-leucine, the title compound wasprepared as a white solid (0.345 g, 67%). MS(ESI): 311.2 (M+H)⁺.

Example 33

Preparation of 2,2′-(N,N′-bis-benzyloxycarbonylglycinyl)carbohydrazide

Following the procedure of Example 29, except substituting N-Cbz-Glycinefor N-Cbz-L-leucine, the title compound was prepared as a white solid(0.719 g, 91%). MS(ESI): 473.1 (M+H)⁺.

Example 34

Preparation of 2,2′-(N,N′-bis-acetyl-L-leucinyl)carbohydrazide

Following the procedure of Example 29, except substitutingN-acetyl-L-leucine for N-Cbz-L-leucine, the title compound was preparedas a white solid (0.153 g, 23%). MS(ESI): 401.3 (M+H)⁺.

Example 35

Preparation of 2,2′-(N,N′-bis- benzyloxycarbonyl-L-alanyl)carbohydrazide

Following the procedure of Example 29, except substitutingN-Cbz-L-alanine for N-Cbz-L-leucine, the title compound was prepared asa white solid (0.762 g, 91%). MS(ESI): 501.1 (M+H)⁺.

Example 36

Preparation of2-(N-benzyloxycarbonyl-L-leucinyl)-2′-[N′-(4-methylpentanoyl)]carbohydrazide

a) N-benzyloxycarbonyl-L-leucine Methyl Ester

To a solution of leucine methyl ester hydrochloride (5.0 g, 27.5 mmol)in 1,4-dioxane (50 nL) was added sodium carbonate (30.3 mL, 2M in water)followed by benzyl chloroformate (4.69 g, 27.5 mmol). The mixturestirred at room temperature for 24 hours when partitioned between ethylacetate and water. The organic layer was collected, dried (MgSO₄),filtered and concentrated to give the title compound as a colorless oil(7.67 g, 100%). ¹HNMR (400 MHz, CDCl₃) δ 7.39 (m, 5H), 5.38 (d, 2H),5.12 (s, 2H), 4.42 (m, 1H), 3.75 (s, 3H), 1.73-1.50 (m, 3H), 0.94 (m,6H).

b) N-benzyloxycarbonyl-L-leucinyl Hydrazide

To a solution of the compound of Example 36(a) (7.67 g, 27.5 mmol) inmethanol (40 mL) was added hydrazine monohydrate (13.5 g, 270 mmol). Thesolution was stirred at room temperature for 24 hours when partitionedbetween water and ethyl acetate. The organic layer was collected, dried(MgSO₄), filtered and concentrated to give the title compound as anoff-white solid (7.67 g, 100%). ¹HNMR (400 MHz, CDCl₃) δ 8.14 (s, 1H),7.38 (m, 5H), 5.64 (d, 1H), 5.09 (dd, 2H), 4.20 (m, 1H), 3.81 (s br,2H), 1.69-1.51 (m, 3H), 0.92 (dd, 6H).

c) 1-benzyloxycarbonylamino-3-methyl-1-(1,3,4-oxadiazol-2-onyl)butane

A solution of the compound of Example 36(b) (1.0 g, 3.58 mmol) inmethylene chloride (12 mL) was added dropwise to a solution of4-nitrophenylchloroformate (0.361 g, 1.79 mmol) in methylene chloride (8mL) at 0° C. The solution warmed to room temperature and stirred for onehour when partitioned between ethyl acetate and water. The organic layerwas washed with aqueous NaHCO₃ then collected, dried (MgSO₄), filteredand concentrated to a residue which was chromatographed (20% ethylacetate/hexane) to give the title compound as a pale yellow solid (0.322g, 59%). ¹HNMR (400 MHz, CDCl₃) δ 9.18 (s, 1H), 7.38 (m, 5H), 5.13 (m,3H), 4.79 (m, 1H), 1.71 (m, 3H), 0.98 (dd, 6H).

d) 4-methylpentanoyl Hydrazide

Following the procedure of Example 36(b) except substituting ethylisocaproate for benzyloxycarbonyl-L-leucinyl methyl ester, the titlecompound was prepared as a white solid (1.8 g, 100%). ¹HNMR (400 MHz,CDCl₃) δ 7.48 (s br, 1H), 3.62 (s br, 2H), 2.13 (t, 2H), 1.51 (m, 3H),0.85 (d, 6H).

e)2-(N-benzyloxycarbonyl-L-leucinyl)-2′-[N′-(4methylpentanoyl)]carbohydrazide

The compounds of Example 36(c) (0.100 g, 0.325 mmol) and Example 36(d)(0.042 g, 0.325 mmol) were combined and dissolved in ethanol (1 mL). Thesolution was brought to reflux for 24 hours then concentrated to a solidyellow residue which was washed with cool methylene chloride to yieldthe title compound as a white solid (0.053 g, 37%). MS (MH⁺): 436.2.

Example 37

Preparation of bis-(Cbz-leucinyl)-1,3-diamino-propan-2-one

Cbz-leucine (500 mg, 1.88 mmol), EDCI (558 mg, 1.88 mmol) was dissolvedin DMF (4.0 ml) with 1,3-diamino-propan-2-ol (85 mg, 0.94 mmol) andHunig's base (0.3 ml, 1.88 mmol) and was stirred at RT overnight. Thereaction was diluted with EtOAc (20 ml) and was extracted with water(2×20 ml). The combined organics were dried with magnesium sulfate,filtered, concentrated in vacuo. The intermediate was then dissolved inacetone (4.0 ml) and Jones reagent (2.0 ml, 1.5 M) was added dropwiseand the reaction was stirred at RT overnight. The excess Jones reagentwas then quenched with isopropanol (1.0 ml), then the reaction wasdiluted with EtOAc (20 ml) and was extracted with water (2×20 ml) toremove the inorganic salts. The combined organics were dried withmagnesium sulfate, filtered, concentrated, and chromatographed (silicagel, 2-5% MeOH/methylene chloride) to give the title compound as a whitesolid (410 mg, 75%). MS(ES) M+H⁺=583, M+Na⁺=605.

Example 38

Preparation of bis-1,3-(4-phenoxy-benzoyl)-diamino-propan-2-one

Following the procedure of Example 37, except substituting“4phenoxy-benzoic acid” for “Cbz-leucine”, the title compound wasprepared: MS(ES) M+H⁺=481, M+Na⁺=503.

Example 39

Preparation of1-(Cbz-leucinyl)-amino-3-(acetyl-leucinyl)-amino-propan-2-one

Following the procedure of Example 37, except substituting “a mixture ofN-Ac-leucine and Cbz-leucine” for “Cbz-leucine”, the title compound wasprepared: MS(ES) M+H⁺=491, M+Na⁺=513.

Example 40

Preparation of 1-(Cbz-leucinyl)-amino-3-(Cbz-glutamyl-t-butylester)-amino-propan-2-one

Following the procedure of Example 37, except substituting “a mixture ofCbz-glutamic acid gamma t-butyl ester and Cbz-leucine” for“Cbz-leucine”, the title compound was prepared: MS(ES) M+H⁺=655.

Example 41

Preparation of1-(Cbz-leucinyl)-amino-3-(Cbz-glutamyl)-amino-propan-2-one

1-(Cbz-leucinyl)-amino-3-(Cbz-glutamyl-t-butyl ester)-amino-propan-2-one(5 mg, 0.007 mmol) was dissolved in a solution of trifluoroacetic acid(0.5 ml) and methylene chloride (0.5 ml), then was stirred at RT for 2h, the reaction was diluted with toluene (10 ml), then was concentratedin vacuo to provide the title compound: MS(ES) M+H⁺=599.

Example 42

Preparation of bis-1,3-(Cbz-leucinyl)-diamino-(S)-butanone-2-one

a) Cbz-leu-ala-bromo Methyl Ketone

Isobutyl chloroformate (1.46 ml, 11.3 mmol) was added dropwise to asolution of Cbz-leu-ala-OH (4.0 g, 11.3 mmol) and N-methyl morpholine(1.24 ml, 11.3 mmol) in THF (40 ml) at 40 degrees C. The reaction wasstirred 15 min, then was filtered, and was washed with ether.Diazomethane (40.1 mmol from 5.9 g of 1-methyl-3-nitro-nitroso-guanidineand 18 ml of 40% KOH in 150 ml of ether) in ether (200 ml) was added andthe reaction was placed in a refrigerator overnight. 30% HBr/AcOH (7 ml)was added dropwise to the crude reaction mixture and was stirred 5minutes. The solution was washed with aqueous citric acid (50 ml×2),saturated aqueous sodium bicarbonate (3×150 ml), then brine (100 ml).The combined organics were dried with magnesium sulfate, filtered, andconcentrated in vacuo to give a solid which was used in the next stepwithout purification, MS(ES) M+H⁺=413 and 415, M+Na⁺=435 and 437.

b) Cbz-leu-leu-azido Methyl Ketone

Cbz-leu-ala-bromo methyl ketone (650 mg, 1.6 mmol) was dissolved in DMF(7 ml), then sodium azide (122 mg, 1.9 mmol) and potassium fluoride (137mg, 2.36 mmol) was added and the reaction was stirred overnight; Thereaction was partitioned between EtOAc and water, then the combinedorganic extracts were dried with magnesium sulfate, filtered,concentrated in vacuo, then chormatographed (2-5% MeOH, methylenechloride, silica gel) to provide the title compound as a white solid(330 mg, 53%), MS(ES) M+Na⁺=398.

c) Cbz-leu-2-amino-4azido-propan-3-ol

Cbz-leu-leu-azido methyl ketone (330 mg, 0.9 mmol) was dissolved in EtOH(5 ml) and sodium borohydride (100 mg, 2.65 mmol) was added at RT andthe reaction was stirred for 15 minutes. The reaction was quenched withwater (10 ml) and was extracted with EtOAc (25 ml). The combined organicextracts were dried with magnesium sulfate, filtered, concentrated togive the title compound without further purification, MS(ES) M+H⁺=378,M+Na⁺=400.

d) Cbz-leu-2-amino-4-amino-propan-3-ol

Cbz-leu-2-amino-4-azido-propan-3-ol (300 mg, 0.8 mmol)was dissolved inMeOH (4 ml) and triethyl amine (0.33 ml, 2.4 mmol), propan-1,3-dithiol(0.35 ml, 3.82 mmol) was added and the reaction was stirred overnight,concentrated in vacuo, then the white solid was washed with hexaneproviding the title compound which was used in the next reaction withoutfurther purification, MS(ES) M+H⁺=352.

e) bis-1,3-(Cbz-leucinyl)-diamino-(S)-butanone-2-ol

Cbz-leu-2-amino-4-amino-propan-3-ol (140 mg, 0.4 mmol) and Cbz-leucine(106 mg, 0.4 mmol) were dissolved in DMF (2 ml) and N-methyl morpholine(0.08 ml, 0.8 mmol) and HBTU (151 mg, 0.4 mmol) and was stirredovernight. The reaction was partitioned between EtOAc and water, thecombined organics were dried with magnesium sulfate, filtered,concentrated to give the title compound, MS(ES) M+H⁺=599, M+Na⁺=621.

f) bis-1,3-(Cbz-leucinyl)-diamino-(S)-butanone-2-one

Bis-1,3-(Cbz-leucinyl)-diamino-(S)-butanone-2-ol (240 mg, 0.4 mmol) wasdissolved in acetone (2 ml). Jones reagent (0.5 ml, 1.5 M) was addeddropwise and the reaction was stirred at RT overnight. The excess Jonesreagent was then quenched with isopropanol (1.0 ml), then the reactionwas diluted with EtOAc (20 ml) and was extracted with water (2×20 ml) toremove the inorganic salts. The combined organics were dried withmagnesium sulfate, filtered, concentrated, and chromatographed (silicagel, 2-5% MeOH/methylene chloride) to give the title compound as a whitesolid (80 mg, 33%). MS(ES) M−H⁺=595.

Example 43

Preparation of1-(Cbz-leucinyl)-amino-3-(Cbz-phenylalanyl)-amino-propan-2-one

Following the procedure of Example 37, except substituting “a mixture ofCbz-phenylalanine and Cbz-leucine” for “Cbz-leucine”, the title compoundwas prepared (70%): MS(ES) M+H⁺=617, M+Na⁺=639.

Example 44

Preparation of1-(Cbz-leucinyl)-amino-3-(Cbz-norleucinyl)-amino-propan-2-one

Following the procedure of Example 37, except substituting “a mixture ofCbz-norleucine and Cbz-leucine” for “Cbz-leucine”, the title compoundwas prepared: MS(ES) M+H⁺=583, M+Na⁺=605.

Example 45

Preparation of 1-(Cbz-leucinyl)-amino-3-(Cbz-norvalinyl)-amino-p -2-one

Following the procedure of Example 37, except substituting “a mixture ofCbz-norvaline and Cbz-leucine” for “Cbz-leucine”, the title compound wasprepared: MS(ES) M+H⁺=569, M+Na⁺=591.

Example 46

Preparation of bis-1,3-(Cbz-leucinyl)-diamino-5-methyl-(S)-hexan-2-one

a) bis-1,3-(Cbz-leucinyl)-diamino-5-methyl-(S)-hexan-2-one

Following the procedure of Example 42(a)-(f), except substituting“Cbz-leu-leu-OH” for “Cbz-leu-ala-OH” the title compound was prepared:MS(ES) M+H⁺=639.

Example 47

Preparation of1-(acetyl-leucinyl)-amino-3-(4-phenoxy-benzoyl)-amino-propan-2-one

Following the procedure of Example 37, except substituting “a mixture ofN-Ac-leucine and 4-phenoxy-benzoic acid” for “Cbz-leucine”, the titlecompound was prepared: MS(ES) M+H⁺=440.

Example 48

Preparation of1-(Cbz-homo-leucinyl)-amino-(Cbz-leucinyl)-3-amino-propan-2-one

Following the procedure of Example 37, except substituting “a mixtureCbz-homo-leucine and Cbz-leucine” for “Cbz-leucine”, the title compoundwas prepared: MS(ES) M+H⁺=597, M+Na⁺=619.

Example 49

Preparation of bis-1,3-(4-(3-chloro-2-cyano-phenoxy)-phenylsulfonamido)-propan-2-one

4-(3-Chloro-2-cyano-phenoxy)-phenyl sulfonyl chloride (1.3 g, 4 mmol,Maybridge) was added to a solution of 1,3-diamino-propan-2-ol (0.18 g, 2mmol) in DMF (10 ml)/N-methyl morpholine (0.44 ml, 4 mmol) and wasstirred 3 h at RT. The reaction was partitioned between water and EtOAcand the combined organics were dried with magnesium sulfate, thenconcentrated in vacuo. The crudebis-1,3-(4(3-chloro-2-cyano-phenoxy)-phenyl sulfonamido)-propan-2-ol(0.28 g, 0.4 mmol) was then dissolved in acetone (1.0 ml) and Jonesreagent (0.44 ml, 1.5 M) was added dropwise, and the reaction wasstirred overnight at RT. The excess Jones reagent was then quenched withisopropanol (1.0 ml), then the reaction was diluted with EtOAc (20 ml)and was extracted with water (2×20 ml) to remove the inorganic salts.The combined organics were dried with magnesium sulfate, filtered,concentrated, and chromatographed (silica gel, 2-5% MeOH/methylenechloride) to give the title compound as a white solid (90 mg, 34%).MS(ES) M+H+=671, M+Na⁺=693.

Example 50

Preparation of bis-1,3-(4-phenoxy-phenyl sulfonamido)-propan-2one

Following the procedure of Example 49, except substituting4-phenoxy-phenyl sulfonyl chloride for4(3-Chloro-2-cyano-phenoxy)-phenyl sulfonyl chloride, the tide compoundwas prepared: MS(ES) M-H⁺=551.

Example 51

Preparation of1-(Cbz-leucinyl)-amino-3-(4-(3chloro-2-cyano-phenoxy)-phenylsulfonamido)-propan-2-one

Cbz-leucine (660 mg, 2.5 mmol), EDCI (480 mg, 2.5 mmol), HOBT (340 mg,2.5 mmol) was dissolved in DMF (10 ml) with 1,3-diamino-propan-2-ol (225mg, 2.5 mmol) and was stirred at RT overnight. N-methyl morpholine (0.41ml, 3.75 mmol) was added followed by 4-(3-Chloro-2-cyano-phenoxy)-phenylsulfonyl chloride (820 mg, 2.5 mmol, Maybridge) was added and thereaction was stirred 3 h at RT. The reaction was partitioned betweenwater and EtOAc and the combined organics were dried with magnesiumsulfate, then concentrated in vacuo. The crude1-(Cbz-leucinyl)-amino-3-(4-(3-chloro-2-cyano-phenoxy)-phenylsulfonamido)-propan-2-ol was then dissolved in acetone (5.0 ml) andJones reagent (3.0 ml, 1.5 M) was added dropwise, and the reaction wasstirred overnight at RT. The excess Jones reagent was then quenched withisopropanol (1.0 ml), then the reaction was diluted with EtOAc (20 ml)and was extracted with water (2×20 ml) to remove the inorganic salts.The combined organics were dried with magnesium sulfate, filtered,concentrated, and chromatographed (silica gel, 2-5% MeOH/methylenechloride), then the product was triturated from methylene chloride togive the title compound as a white solid (26 mg, 2%). MS(ES) M+H⁺=627.

Example 52

Preparation of 1-(Cbz-leucinyl)-amino-3-(tosyl-amino)-propan-2-one

Following the procedure of Example 51, except substituting tosylchloride for 4-(3-Chloro-2-cyano-phenoxy)-phenyl sulfonyl chloride, thetitle compound was prepared: MS(ES) M−H⁺=488.

Example 53

Preparation of1-(Cbz-leucinyl)-amino-3-((4-phenoxy-phenyl)-sulfonamido)-propan-2-one

Following the procedure of Example 51, except substituting4-phenoxy-phenyl-sulfonyl chloride for4-(3-Chloro-2-cyano-phenoxy)-phenyl sulfonyl chloride, the titlecompound was prepared: MS(ES) M+H⁺=568, M+Na⁺=590.

Example 54

Preparation of1-(Cbz-leucinyl)-amino-3-(2-dibenzofuransulfonamido)-propan-2-one

Following the procedure of Example 51, except 2-dibenzofuransulfonylchloride for 4-(3-Chloro-2-cyano-phenoxy)-phenyl sulfonyl chloride, thetitle compound was prepared: MS(ES) M+H⁺=566, M+Na⁺=588.

Example 55

Preparation of1-(Cbz-homo-leucinyl)-amino-3-(2-dibenzofuransulfonamido)-propan-2-one

Following the procedure of Example 51, except 2-dibenzofuransulfonylchloride for 4-(3-Chloro-2-cyano-phenoxy)-phenyl sulfonyl chloride andCbz-homo-leucine for Cbz-leucine, the title compound was prepared:MS(ES) M+Na⁺=602.

Example 56

Preparation of1-(Cbz-leucinyl)-amino-3-(2-dibenzofuransulfonamido)-(S)-butan-2-one

a) 1-(Cbz-leucinyl)-amino-3-(2dibenzofuransulfonamido)-(S)-butan-2-ol

Cbz-leu-2-amino-4-amino-propan-3-ol (150 mg, 0.42 mmol, as described inExample 56(a)-(d)) and 2-dibenzofuransulfonyl chloride were dissolved inDMF (2 ml) and N-methyl morphonline (0.09 ml, 0.84 mmol) and werestirred overnight. The reaction was partitioned between EtOAc and water,the combined organics were dried with magnesium sulfate, filtered,concentrated to give the title compound, MS(ES) M+H⁺=582, M+Na⁺=604.

b) 1-(Cbz-leucinyl)-amino-3-(2-dibenzofuransulfonamido)-(S)-butan-2-one

1-(Cbz-leucinyl)-amino-3-(2-dibenzofuransulfonamido)-(S)-butan-2-ol (240mg, 0.4 mmol) was dissolved in acetone (2 ml). Jones reagent (0.5 ml,1.5 M) was added dropwise and the reaction was stirred at RT overnight.The excess Jones reagent was then quenched with isopropanol (1.0 ml),then the reaction was diluted with EtOAc (20 ml) and was extracted withwater (2×20 ml) to remove the inorganic salts. The combined organicswere dried with magnesium sulfate, filtered, concentrated, andchromatographed (silica gel, 2-5% MeOH/methylene chloride) to give thetide compound as a white solid (70 mg, 29%). MS(ES) M−H⁻=578.

Example 57

Preparation of (S)-Phenylmethyl[1-[[[3-[Benzyloxycarbonyl-leucinyl-amino]-2-oxopropyl]-1-(benzyl)amino]carbonyl]-3-methylbutyl]carbamate

a) 2-hydroxy-3-azido-propanol

Sodium azide (1.7 g, 26 mmol) was added to a solution of glycidol(Aldrich, 1.3 g, 17.5 mmol) in MeOH (45 ml) and water (5 ml) and washeated to 65 degrees C. for 4 h. The reaction was diluted with water (25ml), extracted with EtOAc (2×50 ml); the combined organic layers wereextracted with water (2×50 ml), then brine (50 ml), then were dried withmagnesium sulfate, filtered, concentrated in vacuo, and chromatographed(silica gel, 30% EtOAc/hexanes) to produce a white solid (1.37 g, 67%);MS(ES) M+H⁺=118.4.

b) 2-hydroxy-3-azido-propan-tosylate

Tosyl chloride (2.3 g, 12 mmol) was added to a solution of2-hydroxy-3-azido-propanol (1.17 g, 10 mmol) and triethyl amine (3.6 g,36 mmol) in methylene chloride (50 ml) and was stirred at RT for 4 h.The reaction was diluted with water (20 ml), extracted with EtOAc (2×50ml); the combined organic layers were extracted with pH 7 buffer (2×50ml), then were dried with magnesium sulfate, filtered, concentrated invacuo, and chromatographed (silica gel, 30% EtOAc/hexanes) to produce awhite solid (1.2 g, 44%); MS(ES) M+H⁺=272.2.

c) 2-(Merrifieldpolymer-6-(oxymethylene-tetrahydropyran-acetal)-3-azido-propan-tosylate

c) 2-Hydroxy-3-azido-propan-tosylate (1.2 g, 4.4 mmol) was added to aslurry of Ellman dihydropyran polymer (cf. (3), Scheme 1) (150 mg, 0.3mmol) in ClCH2CH2Cl (25 ml), then pyridinium ptoluenesulfonate (0.84 g,4.4 mmol) and was agitated at 80 degrees C. by gentle bubbling withargon. The polymer was filtered, washed with DMF (2×10 ml), then MeOH(20 ml), then methylene chloride (4×20 ml); IR 2105 cm-1; Magic AngleSpinning 1H NMR: d 8.0, 7.4, 5.0, 3.4.

d) 2-(Merrifieldpolymer-6-(oxymethylene-tetrahydropyran-acetal)-3-azido-propan-N-benzyl-amine

Benzyl amine (0.32 g, 3 mmol) was added to a slurry of 2-(Merrifieldpolymer-&(oxymethylene-tetrahydropyran-acetal)-3-azido-propan-tosylate(500 mg, 1 mmol) in N-methylpyrolidinone (25 ml) and was and wasagitated at 80 degrees C. by gentle bubbling with argon. The polymer wasfiltered, washed with DMF (2×10 ml), then MeOH (20 ml), then methylenechloride (4×20 ml); IR 2105 cm-1; Magic Angle Spinning 1H NMR: d 7.1,4.7, 4.0, 3.8.

e) 2-(Merrifieldpolymer-6-(oxymethylene-tetrahydropyran-acetal)-3-azido-propan-N-benzyl-(Cbz-leucinyl)-amine

Cbz-leucine (0.82 g, 3.0 mmol) was added to a slurry of 2-(Merrifieldpolymer-6-(oxymethylene-tetrahydropyran-acetal)-3-azido-propan-N-benzyl-amine(120 mg, 0.22 mmol) in DMF (10 ml), diisopropyl ethyl amine (1.2 ml, 6mmol) and HATU (Perseptive Biosystems, 2.2 g, 6 mmol) and was shaken atroom temperature overnight. The resin was filtered, washed with DMF(3×10 ml). The above procedure was repeated, and the final resin washedwith MeOH (2×20 ml), then methylene chloride (5×20 ml); IR 2105, 1735,1630 cm-1;. Magic Angle Spinning 1H NMR: d 7.2, 4.7, 4.1.

f) 2-(Merrifieldpolymer-6-(oxymethylene-tetrahydropyran-acetal)-3-amino-propan-N-benzyl-(Cbz-leucinyl)-amine

Propanedithiol (0.5 ml, xx mmol) was added to a slurry of 2-(Merrifieldpolymer-6-(oxymethylene-tetrahydropyran-acetal)-3-azido-propan-N-benzyl-(Cbz-leucinyl)-amine(150 mg, 0.27 mmol) in MeOH (5 ml) and triethylamine (0.5 ml) and wasgently rocked overnight. The resin was filtered, washed with MeOH (2×20ml), then with DMF (1×10 ml), then with methylene chloride (5×20 ml),and was dried in a vacuum oven overnight; IR 1735, 1640, cm⁻¹.

g) 2-(Merrifieldpolymer-6-(oxymethylene-tetrahydropyran-acetal)-3-(Cbz-leucinyl)-amino-propan-N-benzyl-(Cbz-leucinyl)-amine

Cbz-leucine (0.82 g, 3.0 mmol) was added to a slurry 2-(Merrifieldpolymer-6-(oxymethylene-tetrahydropyran-acetal)-3-amino-propan-N-benzyl-(Cbz-leucinyl)-amine(150 mg, 0.27 mmol) in N-methyl pyrollidinone (10 ml), diisopropyl ethylamine (1.2 ml, 6 mmol) and HBTU (2.2 g, 6 mmol) and was shaken at roomtemperature overnight. The resin was filtered, washed with DMF (3×10ml). The above procedure was repeated, and the final resin washed withMeOH (2×10 ml), then methylene chloride (5×20 ml); Magic Angle Spinning1H NMR: d 7.6, 7.4, 5.1, 5.0, 3.4, 0.8.

h) 1-N-benzyl-1-Cbz-leucinyl-amino-3-Cbz-leucinyl-amino-propan-2-ol

2-(Merrifieldpolymer-6(oxymethylene-tetrahydropyran-acetal)-3-(Cbz-leucinyl)-amino-propan-N-benzyl-(Cbz-leucinyl)-amine(150 mg, 0.27 mmol) was shaken as a slurry with 85:5:10TFA/water/methylene chloride (5 ml) for 4 h at RT. The solution wasfiltereed and the filtrate was concentrated in vacuo, thenchromatographed (silica gel, 5% MeOH/methlene chloride) to produce ayellow solid (65 mg, 35%); MS(ES) M+H⁺=675.1.

i) 1-N-benzyl-1-Cbz-leucinyl-amino-3-Cbz-leucinyl-amino-propan-2-one

1-N-benzyl-1-Cbz-leucinyl-amino-3-Cbz-leucinyl-amino-propan-2-ol (65 mg,0.96 mmol) was dissolved in acetone (5 ml) and Jones reagent (2ml,excess) was added dropwise at room temperature and the reaction wasstirred overnight. The excess Jones reagent was then quenched withisopropanol (5 ml) and the reaction was diluted with water (5 ml) andwas extracted with EtOAc (2×20 ml). The combined organic layers wereextracted with water (2×15 ml), then brine (10 ml), then were dried withmagnesium sulfate, filtered, concentrated in vacuo to produce a yellowsolid, which was chromatographed (silica gel, 50% EtOAc/Hexanes ) toproduce a white solid (16.8 mg, 29%); MS(ES) M+H⁺=673.1.

Example 58

Preparation of (S)-Phenylmethyl[1-[[[3-[(2-dibenzofuranylsulfonyl)amino]-2-oxopropyl]-3-(benzyl)amino]carbonyl]-3-methylbutyl]carbamate

a) N-(2-hydroxy-3-N-benzylamino-propyl)phthalimide

N-(2,3-Epoxypropyl)phthalimide (Aldrich, 2.03 g, 10 mmol) was reluxedwith benzyl amine (1.07 g, 10 mmol) in isopropanol (15 ml) for 3 h. Thereaction was cooled to RT, then concentrated in vacuo producing a whitegum, which was triturated with MeOH, then filtered producing a whitesolid (0.48 g, 15%); MS(ES) M+H⁺=311.

b)N-(2-hydroxy-3-(N-benzyl-2-dibenzofuransulfonamide)-propyl)phthalimide

N-(2-hydroxy-3-N-benzylamino-propyl)phthalimide (0.31 g, 1 mmol) wasstirred with 2-dibenzofuransulfonyl chloride (0.27 g, 1 mmol) inN-methyl morpholine (0.8 ml) and DMF (5 ml) overnight. The reaction wasdiluted with water (10 ml), extracted with EtOAc (2×20 ml), the combinedorganic layers were extracted with water (3×20 ml), then brine (20 ml),then were dried with magnesium sulfate, filtered, concentrated in vacuoto produce an oil, which was chromatographed (silica gel, 30%EtOAc/hexanes) to produce a white foam (0.37 g, 69%); MS(ES) M+H⁺=541,MS(ES) M+Na⁺=563, MS(ES− negative) M+HCO₂ ⁻=585.

c) 2-hydroxy-(N-benzyl-2-dibenzofuransulfonamide)-propyl-3-amine

N-(2-hydroxy-3-(N-benzyl-2-dibenzofuransulfonamide)-propyl)phthalimide(0.37 g, 0.69 mmol) was refluxed with hydrazine hydrate (0.34 g, 6.85mmol) in MeOH (7 ml) for 1.5 h. The reaction was cooled to RT, then wasconcentrated in vacuo. The resulting white solid was triturated withMeOH, then filtered to produce the desired product as a white solid(0.27 g, 96%); MS(ES) M+H⁺=411.

d)Cbz-leucinyl-(2-hydroxy-(N-benzyl-2-dibenzofuransulfonamide))-propyl-3-amine

2-hydroxy-(N-benzyl-2-dibenzofuransulfonamide)-propyl-3-amine (0.2 g,0.5 mmol) was stirred with Cbz-leucine (0.13 g, 0.5 mmol) in N-methylmorpholine (0.6 ml) and DMF (2 ml), then HBTU (0.19 g, 0.5 mmol) wasadded and the reaction was stirred overnight at RT. The reaction wasdiluted with water (10 ml), extracted with EtOAc (2×20 ml). A solid thatwas insoluble in both layers was filtered off. The combined organiclayers were extracted with water (2×20 ml), then brine (20 ml), thenwere dried with magnesium sulfate, filtered, concentrated in vacuo toproduce a white solid, which was used in the next reaction withoutfurther purification; MS(ES) M+H⁺=658, MS(ES) M+Na⁺=680.

e) (S)-Phenylmethyl[1-[[[3-[(2-dibenzofuranylsulfonyl)amino]-2-oxopropyl]-3-(benzyl)amino]carbonyl]-3-methylbutyl]carbamate

Cbz-leucinyl-(2-hydroxy-(N-benzyl-2-dibenzofuransulfonamide))-propyl-3-amine(0.16 g, 0.244 mmol) was dissolved in acetone (2 ml). Jones reagent (0.5ml, 1.5 M) was added added and the reaction was stirred overnight. Theexcess Jones reagent was then quenched with isopropanol (1 ml) and thereaction was diluted with water (10 ml) and was extracted with EtOAc(2×20 ml). The combined organic layers were extracted with water (2×20ml), then brine (20 ml), then were dried with magnesium sulfate,filtered, concentrated in vacuo to produce a white solid, which waschromatographed (silica gel, 1:1 EtOAc/hexanes) to produce a white solid(0.14 g, 88%); MS(ES) M−H⁺=654, MS(ES) M+Cl⁺=690, MS(ES) M+HCO₂ ⁻=700.

Example 59

Preparation of (S)-Phenylmethyl[1-[[[3-[(2-dibenzofuranylsulfonyl)amino]-2-oxopropyl]-3-(4-pyridinylmethyl)amino]carbonyl]-3-methylbutyl]carbamate

Following the procedure of Example 58(a)-(e), except substituting“4-pyridyl methyl amine” for “benzylamine” and, the title compound wasprepared; MS(ES) M+H⁺=657.

Example 60

Preparation of1-[[[3-[(2-dibenzofuranylsulfonyl)amino]-2-oxopropyl]-3-(4-pyridinylmethyl)benzamide

Following the procedure of Example 58(a)-(e), except substituting“benzoic acid” for “Cbz-leucine”, the title compound was prepared;MS(ES) M-H=511, MS(ES) M+Cl⁻=547.

Example 61

Preparation of (S)-Phenylmethyl[1-[[[3-[(2-dibenzofuranylsulfonyl)amino]-2-oxopropyl]-1-(4-pyridinylmethyl)amino]carbonyl]-3-methylbutyl]carbamate

Following the procedure of Example 58(a)-(e), except substituting“4-pyridyl methyl amine” for “benzylamine” and “Cbz-leucine and HBTU”for “2-dibenzofuransulfonyl chloride” and “2-dibenzofuransulfonylchloride” for “Cbz-leucine and HBTU”, the title compound was prepared;MS(ES) M+H⁺=657.

Example 62

Preparation of2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-(4-phenoxyphenylsulfonyl)]carbohydrazide

a) N-benzyloxycarbonyl-L-leucine Methyl Ester

To a stirring solution of L-leucine methyl ester hydrochloride (2.0 g,11.0 mmol) in 1,4-dioxane (20 mL) was added Na₂CO₃ (12.1 ml, 2M inwater) followed by benzylchloroformate (1.96 g, 11.5 mmol). The mixturewas stirred at room temperature for 4 h then partitioned between ethylacetate and water. The organic layer was washed with brine, dried(MgSO₄), filtered and concentrated to yield the title compound as acolorless oil (3.1 g, 100%). ¹H NMR (400 MHz, CDCl₃) δ 7.34 (m, 5H),5.27 (d, 1H), 5.12 (s, 2H), 4.41 (s, 2H), 3.75 (s, 3H), 1.65 (m, 3H),0.96 (m, 6H).

b) N-benzyloxycarbonyl-L-leucinylhydrazide

To a stirring solution of the compound of Example 62(a) (3.1 g, 11.0mmol) in 15 mL of methanol was added hydrazide hydrate (5.9 g, 118mmol). The solution was stirred at room temperature for 16 h thenconcentrated to yield the title compound as an off-white solid (3.1 g,100%). MS(ESI): 280.2 (M+H)⁺.

c)(1S)-1-benzyloxycarbonylamino-3-methyl-1-(1,3,4-oxadiazol-2-on-5-yl)butane

To a stirring solution of the compound of Example 62(b) (3.0 g, 10.8mmol) in toluene (50 mL) was added phosgene (56 mL, 1.93M in toluene).The solution was heated at reflux for 4 h then concentrated to yield thetitle compound as a pale yellow foam (3.15 g, 96%). MS(ESI): 306.1(M+H)⁺.

d) 2-[N-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide

To a stirring solution of the compound of Example 62(c) (0.147 g, 0.482mmol) in 2 mL of methanol was added hydrazine hydrate (0.241 g, 4.82mmol). The solution was stirred at room temperature for 24 h thenconcentrated and purified by column chromatography (silica gel,methanol/dichloromethane) to yield the tide compound as a white foam(0.097 g, 60%). MS(ESI): 338.2 (M+H)⁺.

e)2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-(4-phenoxyphenylsulfonyl)]carbohydrazide

To a stirring solution of the compound of Example 62(d) (0.097 g, 0.288mmol) in 2 mL of DMF was added pyridine (0.046 g, 0.576 mmol) followedby 4-phenoxyphenylsulfonylchloride (0.155 g, 0.576 mmol). The solutionwas stirred at room temperature for 16 h then partitioned between ethylacetate and water. The organic layer was washed with brine, dried(MgSO₄), filtered and concentrated. The residue was purified by columnchromatography (silica gel, ethyl acetate/hexane) to yield the titlecompound as a white solid (0.052 g, 32%). MS(ESI): 570.1 (M+H)⁺.

Example 63

Preparation of2-[N-(N-benzyloxycarbonyl-L-alanyl)1-2′-1N-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide

To a stirring solution of the compound of Example 62(d) (0.100 g, 0.297mmol) in 2 mL of DMF was added N-benzyloxycarbonyl-L-alanine (0.070 g,0.312 mmol), 1-hydroxybenzotriazole (0.008 g, 0.059 mmol), and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.060 g,0.312 mmol). After stirring at room temperature for 16 h, the solutionwas poured into 150 mL of water. The precipitate was filtered and washedwith water (150 mL) and dried under high vacuum to yield the titlecompound as a white solid (0.062 g, 39%). MS(ESI): 543.1 (M+H)⁺.

Example 64

Preparation of2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-(4-phenylbenzoyl)]carbohydrazide

Following the procedure of Example 63, except substituting4-phenylbenzoic acid for N-benzyloxycarbonyl-L-alanine, the titlecompound was prepared as a white solid (0.121 g, 53%). MS(ESI): 518.1(M+H)⁺.

Example 65

Preparation of2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-(4-methoxybenzoyl)]carbohydrazide

Following the procedure of Example 63, except substituting4-methoxybenzoic acid for N-benzyloxycarbonyl-L-alanine the titlecompound was prepared as a white solid (0.057 g, 27%). MS(ESI): 472.1(M+H)⁺.

Example 66

Preparation of2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-(4-phenoxybenzoyl)]carbohydrazide

Following the procedure of Example 63, except substituting4-phenoxybenzoic acid for N-benzyloxycarbonyl-L-alanine the titlecompound was prepared as a white solid (0.102 g, 43%). MS(ESI): 534.1(M+H)⁺.

Example 67

Preparation of2-(N-acetyl)-2′-[(N′-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide

To the compound of Example 62(d) (0.100 g, 0.297 mmol) was added aceticanhydride (0.303 g, 2.97 mmol). The solution was stirred at roomtemperature for 16 h then concentrated to an off-white solid which waswashed with dichloromethane to yield the title compound as a white solid(0.086 g, 76%). MS(ESI): 380.1 (M+H)⁺.

Example 68

Preparation of2-[N-(N-acetyl-L-leucinyl)]-2′-[N′-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide

a) 2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide

Following the procedure of Example 62(a)-62(d), except substitutingL-alanine ethyl ester hydrochloride for L-leucine methyl esterhydrochloride in step (a), the title compound was prepared as a paleyellow foam (1.1 g, 3.8 mmol). MS(ESI): 296.2 (M+H)⁺.

b)2-[N-(N-acetyl-L-leucinyl)]-2′-[N′-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide

To a stirring solution of the compound of Example 63(d) (0.150 g, 0.508mmol) in DMF (2 ml) was added N-acetyl-L-leucine (0.092 g, 0.534 mmol),1-hydroxybenzotriazole (0.014 g, 0.102 mmol), and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.102 g,0.534mmol). After stirring at room temperature for 16 h, the solutionwas diluted with ethyl acetate, washed successively with water,saturated aqueous sodium bicarbonate, and brine. The organic layer wasdried (MgSO₄), filtered and concentrated. The residue was purified bycolumn chromatography (silica gel, methanol/dichloromethane) to yieldthe title compound as a white solid (0.028 g, 12%). MS(ESI): 451.1(M+H)⁺.

Example 69

Preparation of2-[N-(N-acetyl-L-alanyl)]-2′-[N-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide

Following the procedure of Example 68(b), except substitutingN-acetyl-L-alanine for N-acetyl-L-leucine and2-[N-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide for2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide, the title compoundwas prepared as a white solid (0.050 g, 25%). MS(ESI): 473.1 (M+Na)⁺.

Example 70

Preparation of2-N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-[4(N,N-dimethylaminomethyl)benzoyl)])carbohydrazide

a) methyl 4-(N,N-dimethylaminomethyl)benzoate

Methyl 4-(bromomethyl)benzoate (2.0 g, 8.73 mmol) was added to asaturated solution of dimethylamine in methanol. After stirring for 25min, the solution was concentrated and the residue was partitionedbetween 1N NaOH and ethyl acetate. The organic layer was washed withsaturated brine, dired (MgSO₄), filtered, and concentrated to providethe title compound as a colorless liquid (1.67 g, 99%). ¹H NMR (250 MHz,CDCl₃) δ 8.00 (d, 2H), 7.39 (d, 2H), 3.91 (s, 3H), 3.47 (d, 2H), 2.25(s, 6H).

b) 4-(N,N-dimethylaminomethyl)benzoic Acid Lithium Salt

The compound of Example 70(a) (1.67 g, 8.6 mmol) was dissolved inTHF/H₂O (1:1) and LiOH.H₂O (0.39 g, 9.3 mmol) was added. The mixture wasstirred at room temperature for 0.5 h, then taken to reflux for 1.5 h.The mixture was concentrated, redissolved in 25 mL of water andreconcentrated to yield a white solid (1.6 g, 100%). ¹H NMR (400 MHz,CD₃OD) δ 7.94 (d, 2H), 7.36 (d, 2H), 3.64 (s, 2H), 2.35 (s, 6H).

c)2-[—(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-[4-(N,N-dimethylaminomethyl)benzoyl)]]carbohydrazide

Following the procedure of Example 68(b), except substituting4-(N,N-dimethylaminomethyl)benzoic acid lithium salt forN-acetyl-L-leucine and2-[N-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide for2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide, the title compoundwas prepared as a pale yellow solid (0.050 g, 17%). MS(ESI): 499.1(M+H)⁺.

Example 71

Preparation of2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-14-hydroxy-[3-(4-morpholinomethyl)]]benzoyl]carbohydrazide

Following the procedure of Example 68(b), except substituting4-hydroxy-3-(4-morpholinomethyl)benzoic acid for N-acetyl-L-leucine and2-[N-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide for2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide, the title compoundwas prepared as a white solid (0.065 g, 26%). MS(ESI): 557.0 (M+H)⁺.

Example 72

Preparation of2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-[4-(N,N-dimethylaminomethyl)benzyloxy]carbonyl-L-leucinyl]carbohydrazide

a) α-isocyanato-L-leucine Methyl Ester

L-leucine methyl ester hydrochloride (25 g, 0.14 mol) was dissolved inmethylene chloride (450 mL), cooled to 0° C., and pyridine (43.5 g, 0.55mol, 44.5 mL) was added, then a 1.93 M solution of phosgene in toluene(0.18 mol, 92.7 mL) was added slowly. After stirring at 0° C. for 2 h,the mixture was poured into 1400 mL of 0.5 N Hcl and 900 mL of ice. Theorganic layer was washed with 1400 mL of 0.5 N Hcl and 900 mL of ice.The aqueous layers were extracted with methylene chloride (450 mL) andthe combined organic layers were washed with 1400 mL of saturated brineand 900 mL of ice, then dried (MgSO₄), filtered and concentrated. Theresidue was distilled (56-58° C.; 0.78 mmHg) to provide the titlecompound as a colorless liquid (20.4 g, 86%). ¹H NMR (250 MHz, CDCl₃) δ4.04 (dd, 1H), 3.82 (s, 3H), 1.92-1.72 (m, 1H), 1.69-1.62 (m, 2H), 0.96(d, 3H), 0.94 (d, 3H).

b) 4-(N,N-dimethylamino)benzyl Alcohol

To a stirring solution of the compound of Example 70(a) (1.63 g, 8.4mmol) in 25 mL of ether, cooled to 0° C., was added dropwise a 1 Msolution of lithium aluminum hydride (8.4 mmol, 8.4 ml). After 5 min,the reaction was quenched by the addition of water (0.33 mL), 15%aqueous NaOH (0.33 mL) and water (1.0 mL). The precipitate was removedby filtration, washed with ether 2 times and the filtrate wasconcentrated to provide the title compound as a colorless oil (1.36 g,98%). ¹H NMR (250 MHz, CDCl₃) δ 7.32 (d, 2H), 7.28 (d, 2H), 4.68 (s,2H), 3.41 (s, 2H), 2.22 (s, 6H).

c) N-[4-(N,N-dimethylaminomethyl)benzyloxycarbonyl]-L-leucine MethylEster

A solution of the compound of Example 72(a) (1.0 g, 5.8 mmol) and thecompound of Example 72(b) in toluene (6 mL) was heated at reflux for 24h. The solution was concentrated and the residue was purified by flashchromatography on 60 g of 230-400 mesh silica gel, eluting with 5%methanol in methylene chloride, to provide the title compound as a paleyellow oil (1.71 g, 87%). ¹H NMR (400 MHz, CDCl₃) δ 7.31 (s, 4H), 5.13(d, 11H), 5.10 (s, 2H), 4.41 (m, 1H), 3.74 (s, 3H), 3.43 (s, 2H), 2.24(s, 6H), 1.70-1.62 (m, 2H), 1.52 (m, 1H), 0.96 (d, 3H), 0.94 (d, 3H).

d) N-[4-(N,N-dimethylaminomethyl)benzyloxycarbonyl]-L-leucine LithiumSalt

Following the procedure of Example 70(b), except substitutingN-[s(N,N-dimethylaminomethyl)benzyloxycarbonyl]-L-leucine methyl esterfor methyl 4-(N,N-dimethylaminomethyl)benzoate, the title compound wasprepared as a white solid (1.57 g, 95%). ¹H NMR (400 MHz, CD₃OD) δ 7.35(d, 2H), 7.30 (d, 2H), 5.06 (dd, 2H), 4.10 (dd, 1H), 3.48 (s, 2H), 2.23(d, 6H), 1.69-1.51 (m, 3H), 0.94 (d, 3H), 0.93 (d, 3H).

e)2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-[4-(N,N-dimethylaminomethyl)benzyloxy]carbonyl-L-leucinyl]carbohydrazide

Following the procedure of Example 68(b), except substitutingN-[4-(N,N-dimethylaminomethyl)benzyloxycarbonyl]-L-leucine lithium saltfor N-acetyl-L-leucine and2-[N-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide for2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide, the title compoundwas prepared as a white solid (0.069 g, 18%). MS(ESI): 642.1 (M+H)⁺.

Example 73

Preparation of2-(N-benzoyl)-2′-[N′-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide

Following the procedure of Example 62(e) except substituting benzoylchloride for 4-phenoxyphenylsulfonylchloride, the title compound wasprepared as a white solid (61 mg, 31%). MS(ESI): 442.1 (M+H)⁺.

Example 74

Preparation of2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-[3-(4-morpholinomethyl)benzoyl]]carbohydrazide

a) methyl 3-(4-morpholinomethyl)benzoate

A solution of morpholine (0.836 g, 9.6 mmol) and methyl3-(bromomethyl)benzoate in THF (5 mL) and DMF (5 mL) was stirred at 50°C. for 3 h. The solution was partitioned between ethyl acetate andwater. The organic layer was washed successively with water, saturatedaqueous NaHCO₃, and brine then dried (MgSO₄), filtered and concentratedto yield a colorless oil (0.872 g, 3.72 mmol). ¹H NMR (400 MHz, CDCl₃) δ7.99 (s, 1H), 7.91 (d, 1H), 7.55 (d, 1H), 7.47 (t, 1H), 3.94 (s, 3H),3.72 (m, 4H), 3.53 (s, 2H), 2.46 (m, 4H).

b) 3-(4-morpholinomethyl)benzoic Acid

To a solution of the compound of Example 74(a) (0.872 g, 3.72 mmol) inTHF (3 mL) and water (3 mL) was added lithium hydroxide monohydrate(0.171 g, 4.08 mmol). After stirring at room temperature for 3 h, thesolution was concentrated. The residue was redissolved in water (5 mL)and 3N HCl was added and the solution was lyophilized to yield a yellowsolid (0.822 g, 3.72 mmol). MS(ESI): 222.0 (M+H)⁺.

c)2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-[3-(4-morpholinomethyl)benzoyl]]carbohydrazide

Following the procedure of Example 68(b), except substituting3-(4-morpholinomethyl)benzoic acid for N-acetyl-L-leucine and2-[N-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide for2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide, the title compoundwas prepared as a white solid (0.056 g, 20%). MS(ESI); 541.0 (M+H)⁺.

Example 75

Preparation of2-[N-(3-benzyloxybenzoyl)]-2′-[N′-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide

a) methyl 3-benzyloxybenzoate

To a suspension of NaH (0.395 g, 9.87 mmol, 60% in mineral oil) in DMF(20 mL) was added methyl 3-hydroxybenzoate (1.0 g, 6.58 mmol). Afterstirring for 15 min at room temperature, benzyl bromide (1.1 g, 6.58mmol) was added. After stirring at room temperature for 3 h, thesolution was partitioned between ethyl acetate and water. The organiclayer was washed with water (2×75 mL), saturated aqueous sodiumbicarbonate, and brine, then dried (MgSO₄), filtered and concentrated toyield an off-white solid (1.013 g, 4.2 mmol). ¹H NMR (400 MHz, CDCl₃) δ7.67 (m, 2H), 7.48-7.34 (m. 6H), 7.19 (m, 1H), 5.12 (s, 2H), 3.95 (s,3H).

b) 3-benzyloxybenzoic Acid

To a solution of the compound of Example 75(a) (0.400 g, 1.65 mmol) inTHF (2 mL) and water (2 mL) was added lithium hydroxide monohydrate(0.076 g, 1.82 mmol). After stirring at reflux for 5 h, the solution waspartitioned between ethyl acetate and 3N HCl. The organic layer waswashed with brine, dried (MgSO₄), filtered and concentrated to yield awhite solid (0.355 g, 1.56 mmol). ¹H NMR (400 MHz, CD₃OD) δ 7.58 (m,2H), 7.36-7.24 (m. 6H), 7.10 (m, 1H), 5.04 (s, 2H).

c)2-[N-(3-benzyloxybenzoyl)]-2′-[N′-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide

Following the procedure of Example 68(b), except substituting3-benzyloxybenzoic acid for N-acetyl-L-leucine and2-[N-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide for2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide, the title compoundwas prepared as a white solid (0.062 g, 25%). MS(ESI): 548.1 (M+H)⁺.

Example 76

Preparation of2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N-[4-[3-(N-N-dimethylamino)-1-propyloxy]benzoyl]]carbohydrazide

a) methyl 4-[3-(N,N-dimethylamino)-1-propyloxy]benzoate

To a solution of methyl 4-hydroxybenzoate (1.0 g, 6.58 mmol),3-dimethylamino-1-propanol (1.01 g, 9.87 mmol), and triphenylphosphine(2.6 g, 9.87 mmol) at 0° C. in THF (20 mL) was added dropwisediisopropyl azodicarboxylate (1.99 g, 9.87 mmol). After stirring for 16h at room temperature the solution was concentrated and the residuepurified by column chromatography (silica gel, methanol/dichloromethane)to yield the title compound as an oily solid (1.25 g, 5.2 mmol).MS(ESI): 238.1 (M+H)⁺.

b) 4-[3-(N,N-dimethylamino)-1-propyloxy]benzoic Acid

Following the procedure of Example 74(b) except substituting methyl4-[3-(N,N-dimethylamino)-1-propyloxy]benzoate for methyl3-(4-morpholinomethyl)benzoate, the title compound was prepared as a tansolid (1.17 g, 5.2 mmol). MS(ESI): 224.1 (M+H)⁺.

c)2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-[4-[3-(N-N-dimethylamino)-1-propyloxy]benzoyl]]carbohydrazide

Following the procedure of Example 68(b), except substituting4[3-(N,N-dimethylamino)-1-propyloxy]benzoic acid for N-acetyl-L-leucineand 2-[N-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide for2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide, the title compoundwas prepared as a white solid (0.060 g, 21%). MS(ESI): 543.1 (M+H)⁺.

Example 77

Preparation of2-[N-(2-benzyloxybenzoyl)]-2′-[N′-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide

Following the procedure of Example 68(b), except substituting2-benzyloxybenzoic acid for N-acetyl-L-leucine and2-[N-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide for2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide, title compound wasprepared as a white solid (0.056 g, 23%). MS(ESI): 548.1 (M+H)⁺.

Example 78

Preparation of2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-[3-(4-pyridylmethoxy)benzoyl]]carbohydrazide

a) methyl 4-pyridinylmethoxybenzoate

Following the procedure of Example 76(a) except substituting methyl3-hydroxybenzoate for methyl 4-hydroxybenzoate and 4-pyridylcarbinol for3-dimethylamino-1-propanol, the title compound was prepared as a yellowsolid (0.599 g, 2.5 mmol). MS(ESI): 244.1 (M+H)⁺.

b) 4-pyridinylmethoxybenzoic Acid

Following the procedure of Example 75(b) except substituting methyl4-pyridylmethoxybenzoate for methyl 3-benzyloxybenzoate the titlecompound was prepared as a yellow solid (0.386 g, 1.69 mmol). ¹H NMR(400 MHz, CD₃OD) δ 8.54 (d, 2H), 7.64 (m, 2H), 7.57 (m, 2H), 7.40 (m,1H), 7.26 (m, 1H), 5.24 (s, 2H).

c)2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-[3-(4-pyridylmethoxy)benzoyl]]carbohydrazide

Following the procedure of Example 68(b), except substituting4-pyridinylmethoxybenzoic acid for N-acetyl-L-leucine and2-[N-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide for2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide, title compound wasprepared as a white solid (0.219 g, 67%). MS(ESI): 549.1 (M+H)⁺.

Example 79

Preparation of2-[N-(4-benzyloxybenzoyl)]-2′-[N′-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide

Following the procedure of Example 75(a)-75(c) except substitutingmethyl 4-hydroxybenzoate for methyl 3-hydroxybenzoate in step (a), thetitle compound was prepared as a white solid (0.160 g, 49%). MS(ESI):548.1 (M+H)⁺.

Example 80

Preparation of2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-(3-benzyloxy-5-methoxy)benzoyl]carbohydrazide

a) methyl 3-hydroxy-5-methoxybenzoate

A suspension of methyl 3,5-dihydroxybenzoate (2.0 g, 11.9 mmol), K₂CO₃(1.6 g, 11.9 mmol), and iodomethane (1.7 g, 11.9 mmol) in acetone (100mL) was stirred at reflux. After stirring for three hours the mixturewas partitioned between ethyl acetate and water. The organic layer waswashed with brine, dried (MgSO₄), filtered and concentrated. The residuewas purified by column chromatography to yield the title compound as awhite solid (0.813 g, 4.4 mmol). ¹H NMR (400 MHz, CDCl₃) δ 7.16 (m, 1H),7.12 (m, 1H), 6.61 (m, 1H), 5.04 (s, 1H), 3.91 (s, 3H) 3.82 (s, 3H).

b) methyl 3-benzyloxy-5-methoxybenzoate

Following the procedure of Example 80(a) except substituting methyl3-hydroxy-5-methoxybenzoate for methyl 3,5-dihydroxybenzoate and benzylbromide for iodomethane, the title compound was prepared as a tan oil(1.2 g, 4.4 mmol). ¹H NMR (400 MHz, CDCl₃) δ 7.45-7.31 (m, 6H), 7.24 (s,1H), 6.76 (m, 1H), 5.09 (s, 2H), 3.95 (s, 3H), 3.84 (s, 3H).

c) 3-benzyloxy-5-methoxybenzoic Acid

Following the procedure of Example 75(b) except substituting methyl3-benzyloxy-5-methoxybenzoate for methyl 3-benzyloxybenzoate, the titlecompound was prepared as an orange solid (1.14 g, 4.4 mmol). ¹HNMR (400MHz, CDCl₃) δ 7.42-7.20 (m, 7H), 6.71 (m, 1H), 5.05 (s, 2H), 3.80 (s,3H).

d)2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-(3-benzyloxy-5-methoxy)benzoyl]carbohydrazide

Following the procedure of Example 68(b), except substituting3-benzyloxy-5-methoxybenzoic acid for N-acetyl-L-leucine and2-[N-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide for2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide, title compound wasprepared as a white solid (0.201 g, 59%). MS(ESI): 578.0 (M+H)⁺.

Example 81

Preparation of2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-(3-benzyloxy-4,5-dimethoxy)benzoyl]carbohydrazide

Following the procedure of Example 68(b), except substituting3-benzyloxy-4,5-dimethoxybenzioc acid for N-acetyl-L-leucine and2-[N-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide for2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide, the title compoundwas prepared as a white solid (0.155 g, 43%). MS(ESI): 607.9 (M+H)⁺.

Example 82

Preparation of2-[N-(N-benzyloxycarbonyl-L-leucinyl)1-2′-[N-(3-benzyloxy-5-ethoxy)benzoyl]carbohydrazide

Following the procedure of Example 80(a)-80(d) except substitutingiodoethane for iodomethane in step (a), the title compound was preparedas a white solid (0.162 g, 46%). MS(ESI): 592.3 (M+H)⁺.

Example 83

Preparation of2-[N-(N-benzyloxycarbonylglycinyl)]-2′-[N′-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide

Following the procedure of Example 68(b), except substitutingN-benzyloxycarbonylglycine for N-acetyl-L-leucine and²-[N-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide for2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide, the title compoundwas prepared as a white solid (0.204 g, 65%). MS(ESI): 529.2 (M+H)⁺.

Example 84

Preparation of2-[N-(3-benzyloxybenzoyl)]-2′-[N′-(N-benzyloxycarbonyl-L-prolinyl)]carbohydrazide

a) 2-[N-(3-benzyloxybenzoyl)]carbohydrazide

Following the procedure of Example 62(b)-62(d) except substitutingmethyl 3-benzyloxybenzoate for N-benzyloxycarbonyl-L-leucine methylester in step (b), the title compound was prepared as an off white solid(0.421 g, 67%). MS(ESI): 301.1 (M+H)⁺.

b)2-[N-(3-benzyloxybenzoyl)]-2′-[N′-(N-benzyloxycarbonyl-L-prolinyl)]carbohydrazide

Following the procedure of Example 68(b), except substitutingN-benzyloxycarbonyl-L-proline for N-acetyl-L-leucine and2-[N-(3-benzyloxybenzoyl)]carbohydrazide for2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide, the tide compoundwas prepared as a white solid (0.219 g, 62%). MS(ESI): 532.0 (M+H)⁺.

Example 85

Preparation of2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-(4-phenylphenylacetyl)]carbohydrazide

Following the procedure of Example 68(b), except substituting4-biphenylacetic acid for N-acetyl-L-leucine and2-[N-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide for2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide, the title compoundwas prepared as a white solid (0.224 g, 71%). MS(ESI): 554.2 (M+Na)⁺.

Example 86

Preparation of(2′S)-2-[N-(3-benzyloxybenzoyl)]-2′-[N′-(N-benzyloxycarbonyl-2-aminobutyryl)]carbohydrazide

Following the procedure of Example 68(b), except substituting(S)-N-benzyloxycarbonyl-2-aminobutyric acid for N-acetyl-L-leucine and2-[N-(3-benzyloxybenzoyl)]carbohydrazide for2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide, the title compoundwas prepared as a white solid (0.244 g, 70%). MS(ESI): 520.3 (M+H)⁺.

Example 87

Preparation of 2,2′-[N,N′-[bis-(4-phenylphenylacetyl)]]carbohydrazide

To a stirring solution of carbohydrazide (0.200 g, 2.22 mmol) in DMF (12mL) was added 4-biphenylacetic acid (1.04 g, 4.89 mmol),1-hydroxybenzotriazole (0.060 g, 0.444 mmol), and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.937 g,4.89 mmol). After stirring at room temperature for 16 h, the solutionwas poured into 150 mL of water. The precipitate was filtered and washedwith water (150 mL) and dried under high vacuum to yield the titlecompound as a white solid (0.977 g, 92%). MS(ESI): 501.1 (M+Na)⁺.

Example 88

Preparation of(2′RS)-2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[2-(4-phenylphenoxy)propionyl]carbohydrazide

Following the procedure of Example 68(b), except substituting2-(4-phenylphenoxy)propionic acid for N-acetyl-L-leucine and2-[N-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide for2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide, the title compoundwas prepared as a white solid (0.183 g, 73%). MS(ESI): 562.3 (M+H)⁺.

Example 89

Preparation of2-[N-(3-benzyloxybenzoyl)]-2′-[N′-(4-methylpentanoyl)]carbohydrazide

Following the procedure of Example 68(b), except substituting4-methylpentanoic acid for N-acetyl-L-leucine and2-[N-(3-benzyloxybenzoyl)]carbohydrazide for2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide, the title compoundwas prepared as a white solid (0.079 g, 30%). MS(ESI): 399.3 (M+H)⁺.

Example 90

Preparation of(2RS-2′RS)-2,2′-[N,N′-[bis-[2-(4-phenylphenyl)-4-methylpentanoyl)]]]carbohydrazide

a) 4-methyl-2-(4-phenylphenyl)pent-4-enoic Acid

To a stirring solution of diisopropylamine (0.537 g, 5.31 mmol) in THF(5.2 mL) at 0° C. was added n-butyllithium (2.1 mL, 5.22 mmol, 2.5M inhexane) dropwise. After stirring for 15 min at 0° C., the mixture wascooled to −78° C. and a solution of 4-biphenylacetic acid (0.500 g, 2.36mmol) in THF (2 mL) was added dropwise. After again warming to 0° C. andcoolling to −78° C., 3-bromo-2-methylpropene (0.485 g, 3.54 mmol) wasadded to the mixture in one portion. After stirring at −78° C. for 1 h,the reaction was quenched with 2 mL of water then concentrated. Theresidue was redissolved in water and extracted with ether (100 mL). Theaqueous layer was acidified (3N HCl) and extracted with ether (3×100mL). The organic layers were combined, dried (MgSO₄), filtered andconcentrated to yield a white solid (0.449 g, 72%). MS(ESI): 265.3(M+H)⁻.

b) 4-methyl-2-(4-phenylphenyl)pentanoic Acid

To a stirring solution of the compound of Example 90(a) (0.449 g, 1.69mmol) in ethyl acetate (25 mL) was added palladium on carbon (0.225 g).After stirring under a balloon of hydrogen for 16 h, the mixture wasfiltered through Celite. The filtrate was concentrated to yield an offwhite solid (0.430 g, 95%). MS(ESI): 267.4 (M+H)⁻.

c)2RS,2′RS)-2,2′-[N,N′-[bis-[2-(4phenylphenyl)-4-methylpentanoyl)]]]carbohydrazide

Following the procedure of Example 87 except substituting4-methyl-2-(4-phenylphenyl)pentanoic acid for 4-biphenylacetic acid, thetitle compound was obtained, after purification by column chromatography(silica gel, methanol/dichloromethane), as a white solid (0.143 g, 33%).MS(ESI): 591.3 (M+H)⁺.

Example 91

Preparation of(2′RS)-2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-[2-(4-phenylphenyl)-4-methylpentanoyl)]]carbohydrazide

Following the procedure of Example 68(b), except4-methyl-2-(4-phenylphenyl)pentanoic acid for N-acetyl-L-leucine and2-[N-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide for2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide, the title compoundwas prepared as a white solid (0.111 g, 42%). MS(ESI): 588.1 (M+H)⁺.

Example 92

Preparation of(2′RS)-2-[N-(3-benzyloxybenzoyl)]-2′-[N′-[2-(4-phenylphenyl)-4-methylpentanoyl)]]carbohydrazide

Following the procedure of Example 68(b), except substituting4-methyl-2-(4-phenylphenyl)pentanoic acid for N-acetyl-L-leucine and2-[N-(3-benzyloxybenzoyl)]carbohydrazide for2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide, the title compoundwas prepared as a white solid (0.195 g, 53%). MS(ESI): 551.1 (M+H)⁺.

Example 93

Preparation of2-[N-(3-benzyloxybenzoyl)]-2′-[N′-(N-benzyloxycarbonyl-N-methyl-L-leucinyl)]carbohydrazide

Following the procedure of Example 68(b), except substitutingN-benzyloxycarbonyl-N-methyl-L-leucine for N-acetyl-L-leucine and2-[N-(3-benzyloxybenzoyl)]carbohydrazide for2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide, the title compoundwas prepared as a white solid (0.341 g, 91%). MS(ESI): 562.2 (M+H)⁺.

Example 94

Preparation of2-[N-(3-benzyloxybenzoyl)]-2′-[N′-[N-(2-pyridinylmethoxycarbonyl)-L-leucinyl]]carbohydrazide

a) N-(2-pyridinylmethoxycarbonyl)-L-leucine Methyl Ester

Following the procedure of Example 72(c), except substituting2-pryidylcarbinol for 4-(N,N-dimethylamino)benzyl alcohol, the titlecompound was prepared as a brown oil (8.06 g, 89%). MS(ESI): 281.2(M+H)⁺.

b) 2-[N-(2-pyridinylylmethoxycarbonyl)-L-leucinyl]carbohydrazide

Following the procedure of Example 62(b)-62(d) except substitutingN-(2-pyridinylmethoxycarbonyl)-L-leucine methyl ester for L-leucinemethyl ester in step (b), the title compound was prepared as a paleyellow foam (0.598 g, 69%). MS(ESI): 339.3 (M+H)⁺.

c)2-[N-(3-benzyloxybenzoyl)]-2′-[N′-[N-(2-pyridinylmethoxycarbonyl)-L-leucinyl]]carbohydrazide

Following the procedure of Example 68(b), except substituting3-benzyloxybenzoic acid for N-acetyl-L-leucine and2-[N-(2-pyridinylylmethoxycarbonyl)-L-leucinyl]carbohydrazide for2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide, the title compoundwas prepared as a white solid (0.057 g, 33%). MS(ESI): 549.2 (M+H)⁺.

Example 95

Preparation of2-[N-[3-(4-pyridylmethoxy)benzoyl]]-2′-[N′-[N-(2-pyridinylmethoxycarbonyl)-L-leucinyl]]carbohydrazide

Following the procedure of Example 68(b), except substituting3-(4-pyridinylmethoxy)benzoic acid for N-acetyl-L-leucine and2-[N-(2-pyridinylylmethoxycarbonyl)-L-leucinyl]carbohydrazide for2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide, the title compoundwas prepared as a yellow solid (0.088 g, 27%). MS(ESI): 550.2 (M+H)⁺.

Example 96

Preparation of(2RS)-2-[N-[2-(4-phenylphenyl)-4-methylpentanoyl)]]-2′-[N′-[N-(2-pyridinylmethoxycarbonyl)-L-leucinyl]]carbohydrazide

Following the procedure of Example 68(b), except4-methyl-2-(4-phenylphenyl)pentanoic acid for N-acetyl-L-leucine and2-[N-(2-pyridinylylmethoxycarbonyl)-L-leucinyl]carbohydrazide for2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide, the title compoundwas prepared as a yellow solid (0.056 g, 24%). MS(ESI): 589.4 (M+H)⁺.

Example 97

Preparation of2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-[2-(4-phenylphenyl)-4-methylpentanoyl)]]carbohydrazide

The title compound was prepared from the compound of Example 91 usingHPLC (Sumipax OA-3100, 4.6×150 mm, 80/20 hexanelethanol, 1.0 mL/min,retention time=5.9 min).

Example 98

Preparation of2-[N-(N-benzyloxycarbonyl-L-leucinyl])-2′-[N′-[2-(4-phenylphenyl)-4-methylpentanoyl)]]carbohydrazide

The tide compound was prepared from the compound of Example 91 usingHPLC (Sumipax OA-3100, 4.6×150 mm, 80/20 hexane/ethanol, 1.0 mL/min,retention time=8.1 min).

Example 99

Preparation of2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-[N-(4-phenylphenyl)-N-(2-methylpropyl)carbamoyl]]carbohydrazide

To a stirring solution of phosgene (0.228 mL, 0.244 mmol, 12.5% solutionin benzene) was added dropwise a solution ofN-(2-methylpropyl)-N-(4-phenylphenyl)amine (0.050 g, 0.222 mmol) andtriethylamine (0.025 g, 0.244 mmol) in dichloromethane (1 mL). Afterstirring at room temperature for 15 min this solution was added dropwiseto a solution of the compound of Example 1(d) (0.083 g, 0.244 mmol) andtriethylamine (0.025 g, 0.244 mmol) in dichloromethane (1 mL) at roomtemperature. After stirring at room temperature for 48 h,N-methylmorpholine (0.022 g, 0.222 mmol) and DMF (2 mL) were added tothe solution and heated at 50° C. for 16 h. The solution was thendiluted with ethyl acetate (5 mL) and washed successively with water,aqueous saturated NaHCO₃, and brine. The organic layer was dried(MgSO₄), filtered and concentrated. The residue was purified by columnchromatography (silica gel, methanol/dichloromethane) to yield the titlecompound as a yellow solid (0.023 g, 18%). MS(ESI): 589.4 (M+H)⁺.

Example 100

Preparation of2-[N-(3-benzyloxybenzoyl)]-2′-[N′-(N-methyl-L-leucinyl)]carbohydrazide

a)2-[N-(3-benzyloxybenzoyl)]-2′-[N′-(N-tert-butoxycarbonyl-N-methyl-L-leucinyl)]carbohydrazide

Following the procedure of Example 68(b), except substitutingN-tert-butoxycarbonyl-N-methyl-L-leucine for N-acetyl-L-leucine and2-[N-(3-benzyloxybenzoyl)]carbohydrazide for2-[N-(N-benzyloxycarbonyl-L-alanyl)]carbohydrazide, the title compoundwas prepared as a white solid (0.183 g, 69%). MS(ESI): 550.4 (M+Na)⁺.

b)2-[N-(3-benzyloxybenzoyl)]-2′-[N′-(N-methyl-L-leucinyl)]carbohydrazide

To a stirring solution of the compound of Example 100(a) (0.100 g, 0.189mmol) in dichloromethane (1 mL) was added trifluoroacetic acid (0.296 g,2.5 mmol). After stirring at room temperature for 15 min, the solutionwas concentrated and the residue was purified by column chromatography(silica gel, methanol/dichloromethane) to yield the title compound as awhite solid (0.055 g, 68%). MS(ESI): 428.4 (M+H)⁺.

Example 101

Preparation of2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N’-(N-methyl-L-leucinyl)]carbohydrazide

Following the procedure of Example 100(a)-100(b), except substituting2-[N-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide for2-[N-(3-benzyloxybenzoyl)]carbohydrazide in step (a), the title compoundwas prepared. MS(ESI); 465.5 (M+H)⁺.

Example 102

Preparation of(1S)-N-[2-[(1-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4--ylcarbonyl]-N′-(4-phenoxyphenylsulfonyl)hydrazide

a) N-benzyloxycarbonyl-L-leucinamide

To a stirring solution of N-benzyloxycarbonyl-L-leucine (4.6 g, 17.3mmol) in THF, cooled to −40° C., was added N-methylmorpholine (3.68 g,36.4 mmol; 4.0 mL) and isobutyl chloroformate (2.37 g, 17.3 mmol; 2.25mL). After stirring for 15 min, ammonia was bubbled through the solutionfor 5 min. The solution was warmed to room temperature, evaporated, andthe residue was dissolved in ethyl acetate, washed with 0.1 N Hcl, andsaturated brine, then dried (MgSO₄), filtered and evaporated to drynessto give the title compound as a white solid (4.58 g, 100%).

b) N-benzyloxycarbonyl-L-leucinethioamide

A solution of the compound of Example 102(a) (4.58 g, 17.3 mmol) andLawesson's reagent (4.21 g, 10.4 mmol) in THF was allowed to stir atroom temperature for 16 h. The solution was concentrated and the residuewas purified by flash chromatography on 230400 mesh silica gel, elutingwith 1:3 EtOAc/hexanes, to provide the title compound as a pale yellowsolid (3.74 g, 77%).

c)(1S)-1-benzyloxycarbonylamino-1-(4-carboethoxythiazol-2-yl)-3-methylbutane

The compound of Example 102(b) (2.20 g, 7.83 mmol) was dissolved inacetone (35 mL), cooled to −10° C., and ethyl bromopyruvate (1.68 g,8.62 mmol, 1.08 mL) was added. After stirring for 1 h, the solution waspoured into methylene chloride/water, then into saturated aqueousNaHCO₃. The aqueous layer was extracted with methylene chloride and thecombined organic layers were washed with saturated brine, dried (MgSO₄),filtered and concentrated. The residue was dissolved in methylenechloride, cooled to −20° C., pyridine (1.36 g, 17.2 mmol, 1.39 mL) andtrifluroracetic anhydride (1.81 g, 8.62 mmol, 1.22 mL) were added. Afterstirring for 1 h, the solution was washed with saturated squeous NaHCO₃and saturated brine, then dired (MgSO₄), filtered, and concentrated. Tgeresidue was purified by flash chromatography on 90 g of 230-400 meshsilica gel, eluting with 1:3 ethyl acetate/hexanes, to provide the titlecompound as a pale yellow oil (2.36 g, 80%). ¹H NMR (400 MHz, CDCl₃) δ8.08 (s, 1H), 7.38 (m, 5H), 5.42 (s, 3H), 5.23-5.07 (m, 3H), 4.42 (q,2H), 2.01-1.62 (m, 3H), 1.41 (t, 3H), 0.99 (d, 6H).

d)(1S)-1-benzyloxycarbonylamino-1-(4-hydrazinocarbonylthiazol-2-yl)-3-methylbutane

The compound of Example 102(c) (2.16 g, 5.73 mmol) was dissolved inethanol (60 mL) and hydrazine hydrate (2.87 g, 57.3 mmol, 2.8 mL) wasadded and the solution was heated at 75° C. for 1 h. The solution wascooled and evaporated to dryness to provide the title compound as a paleyellow foam (2.01 g, 97%). ¹H NMR (400 MHz, CDCl₃) δ 8.35 (bs, 1H), 8.03(s, 1H), 7.37 (m, 5H), 5.29 (d, 1H), 5.14-5.09 (m, 3H), 4.07 (bs, 2H),1.92-1.82 (m, 1H), 1.79-1.66 (m, 2H), 1.00 (d, 6H).

e)(1S)-N-[2-[(1-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-(4-phenoxyphenylsulfonyl)hydrazide

To a stirring solution of the compound of Example 102(d) (275 mg, 0.76mmol) in dichloromethane at room temperature is added pyridine (180 mg,2.28 mmol, 0.2 mL) and 4-phenoxybenzenesulfonyl chloride (408 mg, 1.52mmol). The reaction was stirred for 16 hours and the solvents wereevaporated to a residue which was chromatographed (silica gel, 40% ethylacetate in hexane) to give the title compound as a white solid (0.310g). MS (ESI): 595.6 (M+H⁺).

Example 103

Preparation of(1S)-N-[4-(1-(N-benzyloxycarbonyl-L-leucinylamino)-3-methylbutyl]thiazol-2-ylcarbonyl]-N′-(N-benzyloxycarbonyl-L-leucinyl)hydrazide

a) N-benzyloxycarbonyl-L-leucinyl-L-leucinyl Bromomethylketone

1-Methyl-3-nitro-1-nitrosoguanidine (5.9 g, 40.11 mmol) in ether (200mL) is cooled to 0° C. 40% potassium hydroxide is added slowly and thediazomethane is allowed to collect in the ether solution for 30 minutesat 0° C.

N-benzyloxycarbonyl-L-Leucinyl-L-Leucine (Bachem) (4.0 g, 10.58 mmol) isstirred in tetrahydrofuran at 40° C. N-methylmorpholine (1.07 g, 10.58mmol, 1.16 mL) and isobutyl chloroformate (1.45 g, 10.58 mmol, 1.38 mL)are added. The mixture is stirred at 40° C. for 15 minutes and thenfiltered into a cold flask to remove precipitated salts. To the filteredsolution is added an excess of the previously prepared diazomethanesolution and the mixture is allowed to stand at 0° C. for 16 h. Anexcess of 30% HBr in acetic acid is added at 0° C. and the solution isthen washed successively with 1.0N citric acid, saturated aqueous sodiumbicarbonate (carefully), and brine. The solution is dried over sodiumsulfate, filtered, and evaporated to give the title compound as a whitesolid (4.10 g). ¹H NMR (400 MHz, CDCl₃) δ 7.34 (m, 5H), 6.51 (d, 1H),5.15 (d, 1H), 5.10 (s, 2H), 4.78 (m, 1H), 4.20 (m, 1H), 4.04 (dd, 2H),1.63 (m, 6H), 0.93 (m, 12H).

b)(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-carboethoxythiazol-4-yl)-3′-methylbutyl]-4-methylpentanamide

The compound of Example 103(a) (2.0 g, 4.4 mmol) and ethyl thiooxamate(0.59 g, 4.4 mmol) were refluxed in ethanol for 4 h. The solvent wasevaporated and the residue chromatographed (silica gel, 2.5%methanol/dichloromethane) to give the title compound as a white solid(1.46 g). ¹H NMR (400 MHz; CDCl₃) δ 7.32 (s, 1H), 7.21 (m, 5H), 6.40 (d,1H), 5.13 (dd, 1H), 5.02 (s, 2H), 4.41 (q, 2H), 4.06 (m, 1H), 1.71 (m,2H), 1.47 (m, 4H), 1.33 (t, 3H), 0.73 (m, 12H).

c)(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-hydrazinocarbonylthiazol-4-yl)-3′-methylbutyl]-4-methylpentanamide

Following the procedure of Example 102(d), except substituting(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-carboethoxythiazol-4-yl)-3′-methylbutyl]-4-methylpentanamidefor(1S)-1-benzyloxycarbonylamino-1-(4-carboethoxythiazol-2-yl)-3-methylbutane,the title compound was prepared. MS (ESI): 476.3 (M+H⁺).

d)(1S)-N-[4-[1-(N-benzyloxycarbonyl-L-leucinylamino)-3-methylbutyl]thiazol-2-ylcarbonyl]-N′-(N-benzyloxycarbonyl-L-leucinyl)hydrazide

To a stirring solution of the compound of Example 103(c) (180 mg, 0.38mmol) in dimethylformamide is added N-benzyloxycarbonyl-L-leucine (11 1mg, 0.42 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride (80 mg, 0.42 mmol), and 1-hydroxybenzotriazole (13 mg,0.096 mmol). The reaction mixture is stirred for 16 hours at roomtemperature, filtered, and washed twice with water. The solvent wasevaporated to give the title compound as a white solid. (0.207 g). MS(ESI): 723.9 (M+H⁺).

Example 104

Preparation of(1S)-N-[2-[(1-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-(4-phenylphenylacetyl)hydrazide

Following the procedure of Example 103(d), except substituting(1S)-1-benzyloxycarbonylamino-1-(4-hydrazinocarbonylthiazol-2-yl)-3-methylbutanefor(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-hydrazinocarbonylthiazol-4-yl)-3′-methylbutyl]4-methylpentanamide,and 4-biphenylacetic acid for N-benzyloxycarbonyl-L-leucine, the titlecompound was prepared as a white solid. MS (ESI ): 557.2 (M+H)⁺.

Example 105

Preparation of(1S)-N-[2-[(1-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[3-(4pryidinylmethoxy)benzoyl]hydrazide

a) methyl 3-(4-pyridinylmethoxy)benzoate

To a stirring solution of methyl 3-hydroxybenzoate (1.0 g, 6.58 mmol),4-pyridylcarbinol (1.1 g, 9.87 mmol), and triphenylphosphine (2.6 g,9.87 mmol) in THF (25 mL) at 0° C. was added diisopropylazodicarboxylate (2.0 g, 9.87 mmol) dropwise. After stirring at roomtemperature for 16 h, the solution was concentrated and purified bycolumn chromatography (silica gel, ethyl acetate/hexane) to yield thetitle compound as a white solid (0.599 g, 37%). MS(ESI): 244.1 (M+H)⁺.

b) 3-(4-pyridinyllmethoxy)benzoic Acid

To a stirring solution of the compound of Example 105(a) (0.599 g, 2.47mmol) in THF/H₂O (1:1, 10 mL) was added lithium hydroxide monohydrate(0.113 g, 2.71 mmol). After stirring at reflux for 3.5 h, 1.1 eq of 1NHCl was added and the mixture poured into water. The mixture wasextracted with ethyl acetate (2×100 mL). The organic layers werecombined, washed with brine, dried (MgSO₄), filtered and concentrated toyield the title compound as a yellow solid (0.386 g, 68%). ¹H NMR (400MHz, CD₃OD) δ 8.54 (d, 2H), 7.64 (m, 2H), 7.57 (m, 2H), 7.40 (m, 1H),7.26 (m, 1H), 5.24 (s, 2H).

c)(1S)-N-[2-[(1-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[3-(4-pryidinylmethoxy)benzoyl]hydrazide

Following the procedure of Example 103(d), except substituting(1S)-1-benzyloxycarbonylamino-1-(4-hydrazinocarbonylthiazol-2-yl)-3-methylbutanefor(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-hydrazinocarbonylthiazol-4-yl)-3′-methylbutyl]-4-methylpentanamide,and 3-(4-pyridinylmethoxy)benzoic acid forN-benzyloxycarbonyl-L-leucine, the title compound was prepared as awhite solid. MS (ESI): 574.2 (M+H)⁺.

Example 106

Preparation ofN-[2-(2-chlorophenoxymethyl)thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

a) α-isocyanato-L-leucine Methyl Ester

L-leucine methyl ester hydrochloride (25 g, 0.14 mol) was dissolved inmethylene chloride (450 mL), cooled to 0° C., and pyridine (43.5 g, 0.55mol, 44.5 mL) was added, then a 1.93 M solution of phosgene in toluene(0.18 mol, 92.7 mL) was added slowly. After stirring at 0° C. for 2 h,the mixture was poured into 0.5 N HCl (1400 mL) and ice (900 mL). Theorganic layer was washed with 0.5 N HCl (1400 mL) and ice (900 mL). Theaqueous layers were extracted with methylene chloride (450 mL) and thecombined organic layers were washed with saturated brine (1400 mL) andice (900 mL), then dried (MgSO₄), filtered and concentrated. The residuewas distilled (56-58° C.; 0.78 mmHg) to provide the title compound as acolorless liquid (20.4 g, 86%). ¹H NMR (250 MHz, CDCl₃) δ 4.04 (dd, 1H),3.82 (s, 3H), 1.92-1.72 (m, 1H), 1.69-1.62 (m, 2H), 0.96 (d, 3H), 0.94(d, 3H).

b) N-(4-pyridinylmethoxycarbonyl)-L-leucine Methyl Ester

A solution of the compound of Example 106(a) (5.10 g, 29.8 mmol) and4-pyridylcarbinol (3.25 g, 29.8 mmol) in toluene (30 mL) was heated atreflux for 24 h. The solution was concentrated and the residue waspurified by flash chromatography on 250 g of 230-400 mesh silica gel,eluting with 3:1 ethyl acetate/hexanes, to give the title compound (7.86g, 94%). ¹H NMR (250 MHz, CDCl₃) δ 8.59 (d, 2H), 7.24 (d, 2H), 5.33 (d,1H), 5.13 (s, 3H), 4.40 (dt, 1H), 3.75 (s, 3H), 1.81-1.51 (m, 3H), 0.96(d, 3H), 0.95 (d, 3H).

c) N-(4pyridinylmethoxycarbonyl)-L-leucine

To a stirring solution the compound of Example 106(b) (1.98 g, 7.06mmol) in THF (7 mL) was added 7 mL of water followed by LiOH.H₂O (325mg, 7.76 mmol). The mixture was stirred for 30 minutes and thenconcentrated. The residue was redissolved in water (10 mL) and 3 NHClwas added (2.6 mL). The solution was lyophilized to yield a white solid(2.015 g, 6.44 mmol). MS (ESI): 267.2 (M+H)⁺.

d) N-[2-(2-chlorophenoxymethyl)thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 102(d), except substituting ethyl2-(2-chlorophenoxymethyl)thiazole-4-carboxylate for(1S)-1-benzyloxycarbonylamino-1-(4-carboethoxythiazol-2-yl)-3-methylbutane,the title compound was prepared. MS (ESI): 284.1 (M+H)⁺.

e)N-[2-(2-chlorophenoxymethyl)thiazol-14ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 103(d), except substitutingN-[2-(2-chlorophenoxymethyl)thiazol-1ylcarbonyl]hydrazide for(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-hydrazinocarbonylthiazol-4-yl)-3′-methylbutyl]4-methylpentanamide,and N-(4-pyridinylmethoxycarbonyl)-L-leucine forN-benzyloxycarbonyl-L-leucine, the title compound was prepared as awhite solid. MS (ESI): 532.1 (M+H)⁺.

Example 107

Preparation ofN-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]-N′-[2-[4-(1,2,3-thiadiazol-4-yl)phenyl]thiazol-4-ylcarbonyl]hydrazide

a) N-[2-[4-(1,2,3-thiadiazol-4-yl)]thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 102(d), except substituting ethyl2-[4-(1,2,3-thiadiazol-4-yl)]thiazole-4-carboxylate for(1S)-1-benzyloxycarbonylamino-1-(4-carboethoxythiazol-2-yl)-3-methylbutane,the tide compound was prepared as a white solid. MS (ESI): 304.1 (M+H)⁺.

b)N-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]-N′-[2-[4-(1,2,3-thiadiazol-4-yl)phenyl]thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 103(d), except substitutingN-[2-(4-(1,2,3-thiadiazol-4-yl)]thiazol-4-ylcarbonyl]hydrazide for(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-hydrazinocarbonylthiazol-4-yl)-3′-methylbutyl]4-methylpentanamide,and N-(4pyridinylmethoxycarbonyl)-L-leucine forN-benzyloxycarbonyl-Leucine, the tide compound was prepared as a whitesolid. MS (ESI): 552.1 (M+H)⁺.

Example 108

Preparation ofN-[2-[3-(4chlorophenylsulfonylmethyl)thien-2-yl]thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

a)N-[2-[3-(4chlorophenylsulfonylmethyl)thien-2-yl]thiazol-4-carbonyl]hydrazide

Following the procedure of Example 102(d), except substituting 2-[3-(4chlorophenylsulfonylmethyl)thien-2-yl]thiazole-4-carboxylate for(1S)-1-benzyloxycarbonylamino-1-(4-carboethoxythiazol-2-yl)-3-methylbutane,the title compound was prepared as a white solid. MS (ESI): 414.1(M+H)⁺.

b)N-[2-[3-(4-chlorophenylsulfonylmethyl)thien-2-yl]thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 103(d), except substitutingN-(2-[3-(4-chlorophenylsulfonylmethyl)thien-2-yl]thiazol-4-carbonyl]hydrazidefor(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-hydrazinocarbonylthiazol-4-yl)-3′-methylbutyl]-4-methylpentanamide,and N-(4-pyridinylmethoxycarbonyl)-L-leucine forN-benzyloxycarbonyl-L-leucine, the title compound was prepared as awhite. MS (ESI): 664.0 (M+H)⁺.

Example 109

Preparation of(1S,2′RS)-N-[2-[(1-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[2′-(4-phenylphenylacetyl)-4-methylpentanoyl]hydrazide

a) 4-methyl-2-(4-phenylphenyl)pent-4-enoic Acid

To a stirring solution of diisopropylamine (0.537 g, 5.31 mmol) in THF(5.2 mL) at 0° C. was added n-butyllithium (2.1 mL, 5.22 mmol, 2.5M inhexane) dropwise. After stirring for 15 min at 0° C., the mixture wascooled to −78° C. and a solution of 4-biphenylacetic acid (0.500 g, 2.36mmol) in THF (2 mL) was added dropwise. After again warming to 0° C. andcoolling to −78° C., 3-bromo-2-methylpropene (0.485 g, 3.54 mmol) wasadded to the mixture in one portion. After stirring at −78° C. for 1 h,the reaction was quenched with 2 mL of water then concentrated. Theresidue was redissolved in water and extracted with ether (100 mL). Theaqueous layer was acidified (3N HCl) and extracted with ether (3×100mL). The organic layers were combined, dried (MgSO₄), filtered andconcentrated to yield a white solid (0.449 g, 72%). MS(ESI)-265.3(M+H)—.

b) 4-methyl-2-(4-phenylphenyl)pentanoic Acid

To a stirring solution of the compound of Example 109(a) (0.449 g, 1.69mmol) in ethyl acetate (25 mL) was added palladium on carbon (0.225 g).After stirring under a balloon of hydrogen for 16 h, the mixture wasfiltered through Celite. The filtrate was concentrated to yield an offwhite solid (0.430 g, 95%). MS(ESI): 267.4 (M+H)—.

c)(1S,2′RS)-N-[2-[(1-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[2′-(4-phenylphenylacetyl)-4-methylpentanoyl]hydrazide

Following the procedure of Example 101(d), except substituting(1S)-1-benzyloxycarbonylamino-1-(4hydrazinocarbonylthiazol-2-yl)-3-methylbutanefor(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-hydrazinocarbonylthiazol-4-yl)-3′-methylbutyl]-4-methylpentanamide,and 4-methyl-2-(4-phenylphenyl)pentanoic acid forN-benzyloxycarbonyl-L-leucine, the tide compound was prepared as a whitesolid. MS (ESI): 613.2 (M+H)⁺.

Example 110

Preparation ofN-[2-(3-benzyloxyphenyl)thiazol-4-ylcarbonyl]-N′-[N-(2-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

a) methyl 3-benzyloxybenzoate

To a suspension of NaH (0.395 g, 9.87 mmol, 60% in mineral oil) in DMF(20 mL) was added methyl 3-hydroxybenzoate (1.0 g, 6.58 mmol). Afterstirring for 15 min at room temperature, benzyl bromide (1.1 g, 6.58mmol) was added. After stirring at room temperature for 3 h, thesolution was partitioned between ethyl acetate and water. The organiclayer was washed with water (2×75 mL), saturated aqueous sodiumbicarbonate, and brine, then dried (MgSO₄), filtered and concentrated toyield an off-white solid (1.013 g, 4.2 mmol). ¹H NMR (400 MHz, CDCl₃) δ7.67 (m, 2H), 7.48-7.34 (m. 6H), 7.19 (m, 1H), 5.12 (s, 2H), 3.95 (s,3H).

b) 3-benzyloxybenzamide

To a suspension of ammonium hydrochloride (1.070 g, 0.02 mmol) in 20 mLof toluene at 5° C., was slowloy added a 2M solution (10 mL) oftrimethylaluminium in toluene. After the addition was complete, thereaction mixture was allowed to warm at room temperature and was stirredfor 2 hours until gas evolution has ceased.

To a stirring solution of the compound of Example 110(a) (605 mg, 2.49mmol) in toluene was added a 0.67 M solution of MeAlClNH₂ (11 mL, 7.49mmol) in toluene. The reaction mixture was allowed to stir overnight atreflux. The reaction was quenched with 5% HCl, the organic layer wasseparated and the aqueous layer extracted three times with ethylacetate. The organic extracts were combined, dried over MgSO₄, filteredand concentrated to afford the title compound as a white solid (409 mg,72%). MS (ESI): 228.1 (M+H)⁺.

c) N-[2-(3-benzyloxyphenyl)thiazolylcarbonyl]hydrazide

Following the procedure of Example 102(b)-102(d), except substituting3-benzyloxybenzamide for N-benzyloxycarbonyl-L-leucinamide in step (b),the title compound was prepared as a white solid. MS (ESI): 326.2(M+H)⁺.

d) N-(2-pyridinylmethoxycarbonyl)-L-leucine

Followong the procedure of Example 106(a)-106(c), except substituting2-pyridylcarbinol for 4pyridylcarbinol in step (b), the title compoundwas prepared. ¹H NMR (400 MHz, CD₃OD) δ 8.50 (d, 1H), 7.86 (dt, 1H),7.51 (d, 1H), 7.36 (dd, 1H), 5.20 (d, 1H), 5.16 (d, 1H), 4.19 (t, 1H),1.78-1.72 (m, 1H), 1.62 (t, 2H), 0.97 (d, 3H), 0.94 (d, 3H).

e)N-[2-(3-benzyloxyphenyl)thiazolylcarbonyl]-N′-[N-(2-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 103(d), except substitutingN-[2-(3-benzyloxyphenyl)thiazol-4-ylcarbonyl]hydrazide for(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-hydrazinocarbonylthiazol-4-yl)-3′-methylbutyl]4methylpentanamide,and N-(2-pyridinylmethoxycarbonyl)-L-leucine forN-benzyloxycarbonyl-L-leucine, the title compound was prepared as awhite solid (106 mg, 0.184 mmol). MS (ESI): 574.2 (M+H)⁺.

Example 111

Preparation of(1RS)-N-[2-[1-(4-phenylphenyl-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

a) N-[2-[1-(4-phenylphenyl)-3-methylbutyl]thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 102(a)-102(d), except substituting4-methyl-2-(4-phenylphenyl)pentanoic acid forN-benzyloxycarbonyl-L-leucine in step (a), the title compound wasprepared as a white solid. MS (ESI): 366.3 (M+H)⁺.

b)(IRS)-N-[2-[1-(4-phenylphenyl)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 103(d), except substitutingN-[2-[1-(4-phenylphenyl)-3-methylbutyl]thiazol-4-ylcarbonyl]hydrazidefor(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-hydrazinocarbonylthiazol-4-yl)-3′-methylbutyl]-4-methylpentanamide,and N-(4-pyridinylmethoxycarbonyl)-L-leucine forN-benzyloxycarbonyl-L-leucine, the title compound was prepared as awhite solid. MS (ESI): 614.3 (M+H)⁺.

Example 112

Preparation ofN-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

a) ethyl 2-aminothiazole-4-carboxylate Hydrobromide

To a stirring suspension of thiourea (6.0 g, 78.8 mmol) in ethanol (80mL) was added ethyl bromopyruvate (15.4 g, 78.8 mmol). The resultingsolution was heated at 45° C. for 23 h. The solution was cooled at 0° C.for 24 h, and the crystals were collected by filtration and washed withcold ethanol to provide the title compound (15.8 g, 79%). ¹H NMR (400MHz, CD₃OD) δ 7.70 (s, 1H), 4.41 (q, 2H), 1.38 (t, 3H).

b) ethyl 2-bromothiazole-4-carboxylate

To a stirring suspension of the compound of Example 112(a) (12.15 g, 48mmol) in 16% aqueous HBr (150 mL), cooled to 0° C., was added drropwis asolution of sodium nitrite (3.44 g, 49.8 mmol) in water (6 mL). Afterstirring for 35 min, copper (I) bromide (7.83 g, 54.6 mmol) and 16%aqueous HBr (60 mL) were added and the mixture was heated at 70° C. for1 h. The mixture was filtered and the filtrate was saturated with NaClthen extracted with ethyl acetate (2×170 mL). The combined extracts weredried (MgSO₄), filtered and evaporated to dryness. The residue wascombined with combined with the solid collected in the first filtration,heated at reflux in ethanol (500 mL) for 5 min, then filtered. To thefiltrate was added 1.5 mL of 48% aqueous HBr and the solution was heatedat reflux for 16 h, then concentrated. The residue was partitionedbetween saturated aqueous NaHCO₃ and ethyl acetate. The organic layerwas washed with saturated brine, dried (MgSO₄), decolorized withcharcoal, filtered and concentrated to provide the title compound as apale yellow solid (7.46 g, 75%). MS (ESI): 236.0 (M+H)⁺.

c) 2-benzyloxybromobenzene

To a stirring solution of 2-bromophenol (10.0 g, 57.8 mmol), and benzylbromide (9.9 g, 57.8 mmol) in acetone (150 mL) was added K₂CO₃ (12.0 g,86.7 mmol). After stirring at reflux for 4 h, the mixture waspartitioned between ethyl acetate and water. The organic layer waswashed with brine, dried (MgSO₄), filtered and concentrated. The residuewas purified by column chromatography (silica gel, ethyl acetate/hexane)to yield the title compound as a colorless oil (15.2 g, 57.8 mmol).¹HNMR (400 MHz, CDCl₃) δ 7.62 (m, 1H), 7.54 (m, 2H), 7.45 (m, 2H), 7.37(m, 1H), 7.28 (m, 1H), 6.98 (m, 1H), 6.91 (m, 1H), 5.17 (s, 2H).

d) 2-benzyloxyphenylboronic Acid

To a stirring solution of the compound of Example 112(c) (15.2 g, 57.8mmol) in THF (100 mL) at −78° C. was added dropwise n-BuLi (23.1 mL,2.5M in hexane, 57.8 mmol). The mixture stirred at −78° C. for 25 minwhen added via cannulation to a stirring solution of triisopropylborate(54.4 g, 289 mmol) in THF (100 mL) at −78° C. After warming to roomtemperature and stirring for 3 h, the mixture was poured into 3N HCl(100 mL) and extracted with ethyl acetate (3×200 mL). The organic layerswere combined, washed successively with water and brine, dried (MgSO₄),filtered and concentrated. The residue was purified by columnchromatography (silica gel, ethyl acetate/hexane) to yield the titlecompound as a pale yellow solid (6.9 g, 30.3 mmol). ¹HNMR (400 MHz,CDCl₃) δ 7.90 (d, 1H), 7.42 (m, 6H), 7.07 (t, 1H), 7.02 (d, 1H), 6.05(s, 2H), 5.16 (s, 2H).

e) ethyl 2-(2-benzyloxyphenyl)thiazole-4-carboxylate

To a stirring solution of the compound of Example 112(b) (4.0 g, 16.9mmol), the compound of Example 72(d) (4.29 g, 18.8 mmol),tetrakis(triphenylphosphine)palladium(0) (0.65 g, 0.57 mmol) indimethoxyethane (60 mL) was added cesium fluoride (8.58 g, 56.5 mmol)and the mixture was heated at 85° C. for 16 h.Tetrakis(triphenylphosphine)palladium(0) (0.65 g, 057 mmol) was addedand heating at 85° C. was continued for 5 h. The mixture was dilutedwith water (60 mL) and extracted with ethyl acetate (2×120 mL). Thecombined extracts were washed with saturated aqueous NaHCO₃ andsaturated brine, dried (MgSO₄), filtered and concentrated. The residuewas purified by flash chromatography on 180 g of 230-400 mesh silicagel, eluting with 15% ethyl acetate in hexanes, to provide the titlecompound as a white solid (3.22 g, 56%). MS (ESI): 340.3 (M+H)⁺.

f) 2-(2-benzyloxyphenyl)thiazol-4-ylcarbonylhydrazide

Following the procedure of Example 102(d), except substituting ethyl2-(2-benzyloxyphenyl)thiazole-4-carboxylate for(1S)-1-benzyloxycarbonylamino-1-(4-carboethoxythiazol-2-yl)-3-methylbutane,the title compound was prepared as a white solid. MS (ESI): 326.2(M+H)⁺.

g)N-[2-(2-benzyloxyphenyl)thiazol-4ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 103(d), except substituting2-(2-benzyloxyphenyl)thiazole-4-ylcarbonylhydrazide for(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-hydrazinocarbonylthiazol-4-yl)-3′-methylbutyl]-4-methylpentanamide,and N-(4-pyridinylmethoxycarbonyl)-L-leucine forN-benzyloxycarbonyl-L-leucine, the title compound was prepared as awhite solid. MS (ESI): 574.3 (M+H)⁺.

Example 113

Preparation ofN-[2-[N-methyl-N-(4-phenylphenyl)amino]thiazol-4-ylcarbonyl]-N′-N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

a) N-(4-phenylphenyl)-2-methylpropionamide

To a stirring solution of 4-aminobiphenyl (9.53 g, 56.3 mmol) andtriethylamine (5.70 g, 56.3 mmol, 7.85 mL) in methylene chloride (60mL), cooled to 0° C., was added slowly isobutyryl chloride (6.0 g, 56.3mmol, 5.90 mL). After stirring at 0° C. for 1 h, the mixture was dilutedwith methylene chloride (120 mL) and washed with 1N NaOH and saturatedbrine, then dreid (MgSO₄), filtered and concentrated. The residue waswashed with ether and dried to provide the title compound as a paleyellow crystalline solid (9.83 g, 73%). ¹HNMR (400 MHz, CDCl₃/CD₃OD) δ7.58 (d, 2H), 7.50 (m, 4H), 7.40-7.25 (m, 3H), 2.55-2.49 (m, 1H), 1.18(d, 6H).

b) N-(4-phenylphenyl)-N-(2-methyl-1-propyl)amine

To a stirring solution of lithium aluminum hydride (58.6 mmol) in THF(58.6 mmol), cooled to 0° C., was added slowly over 10 min a solution ofthe compound of Example 73(a) (9.35 g, 39.0 mmol) in THF (170 nL). Afterthe addition was complete, the ice bath was removed and the solution washeated at 55° C. for 30 min. The mixture was cooled to 0° C. and water(2.22 mL) was slowly added, followed by 15% aqueous NaOH (2.22 mL) andwater (6.67 nL). The precipitate was removed by filtration and washedwith ether 4 times. The filtrate was evaporated to dryness to proveidethe title compound as a pale yellow solid (8.34 g, 97%). MS (ESI): 226.2(M+H)⁺.

c) N-(4-phenylphenyl)-N-(2-methyl-1-propyl)thiourea

To a stirring solution of thiophosgene (98.9 mg, 2.6 mmol, 198 uL) inmethylene chloride (6.5 mL), cooled to 0° C., was added dropwise asolution of the compound of Example 73(b) (540.7 mg, 2.0 mmol) inmethylene chloride (1 mL). After stirring for 2 h. ammonia-satruatedmethanol (20 mL) was added and the solution was stirred at roomtemperatur for 2 h. The solution was concentrated and the residue waspartitioned between ethyl acetate and 1N HCl. The organic layer waswashed with 1N HCl twice, then with saturated brine, then dried (MgSO₄),filtered and concentrated. The residue was purified by flashchromatography on 10 g of 230-400 mesh silica gel, eluting with 1:3ethyl acetate/hexanes, to provide the title compound as a pale yellowsolid (470 mg, 83%). MS (ESI): 285.3 (M+H)⁺.

d) ethyl2-[N-(4-phenylphenyl)-N-(2-methyl-1-propyl)amino]thiazole-4-carboxylate

A solution of the compound of Example 113(c) (184.6 mg, 0.65 mmol) andethyl bromopyruvate (126.6 mg, 0.65 mmol, 81.5 uL) in ethanol (2.5 mL)was heated at reflux fo 5 min, then concentrated. The residue waspartitioned between ethyl acetate and saturated aqueous NaHCO₃. Theaqueous layer was extracted with ethyl acetate and the combined organiclayers were washed with saturated brine, dried (MgSO₄), filtered andconcentrated. The residue was passed through a plug of 230-400 meshsilica gel, eluting with 12% ethyl acetate in hexanes, to provide thetitle compound as a pale yellow oil (230 mg, 93%). MS (ESI): 381.4(M+H)⁺.

e)N-[2-[N-(4-phenylphenyl)-N-(2-methyl-1-propyl)amino]thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 102(d), except substituting ethyl2-[N-(4-phenylphenyl)-N-(2-methyl-1-propyl)amino]thiazole-4-carboxylatefor(1S)-1-benzyloxycarbonylamino-1-(4-carboethoxythiazol-2-yl)-3-methylbutane,the title compound was prepared as a white solid. MS (ESI): 367.3(M+H)⁺.

f)N-[2-[N-methyl-N-(4-phenylphenyl)amino]thiazolylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 103(d), except substitutingN-[2-[N-(4-phenylphenyl)-N-(2-methyl-1-propyl)amino]thiazol-4-ylcarbonyl]hydrazidefor(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-hydrazinocarbonylthiazol-4-yl)-3′-methylbutyl]4-methylpentanamide,and N-(4-pyridinylmethoxycarbonyl)-L-leucine forN-benzyloxycarbonyl-L-leucine, the title compound was prepared as awhite solid. MS (ESI): 615.3 (M+H)⁺.

Example 114

Preparation ofN-(N-benzyloxycarbonyl-L-leucinyl)-N′-[2-(4phenylbenzyl)thiazol-4-ylcarbonyl]hydrazide

a) N-[2-(4-phenylbenzyl)thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 102(a)-102(d), except substituting4-biphenylacetic acid for N-benzyloxycarbonyl-L-leucine in step (a), thetitle compound was prepared as a white solid. MS (ESI): 310.3 (M+H)⁺.

b)N-(N-benzyloxycarbonyl-L-leucinyl)N′-[2-(4-phenylbenzyl)thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 103(d), except substitutingN-[2-(4-phenylbenzyl)thiazol-4-ylcarbonyl]hydrazide for(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-hydrazinocarbonylthiazol-4-yl)-3′-methylbutyl]-4-methylpentanamide,the title compound was prepared as a white solid (20 mg, 0.035 mmol). MS(ESI): 557.4 (M+H)⁺.

Example 115

Preparation ofN-[2-(4phenylphenylbenzyl)thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 103(d), except substitutingN-[2-(4-phenylbenzyl)thiazol-4-ylcarbonyl]hydrazide for(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-hydrazinocarbonylthiazol-yl)-3′-methylbutyl]-4-methylpentanamide,and N-(4-pyridinylmethoxycarbonyl)-L-leucine forN-benzyloxycarbonyl-L-leucine, the title compound was prepared as ayellow solid (30 mg, 0.053 mmol). MS (ESI): 558.2 (M+H)⁺.

Example 116

Preparation ofN-(N-benzyloxycarbonyl-L-leucinyl)-N′-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]hydrazide

a)N-[2-[N-phenyl-N-(2-methyl-1-propyl)amino]thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 113(a)-113(e), except substitutinganiline for 4-aminobiphenyl in step (a), the title compound was preparedas an orang-pink solid (276 mg, 0.950 mmol). MS (ESI): 291.3 (M+H)⁺.

b)N-(N-benzyloxycarbonyl-L-leucinyl)-N′-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 103(d), except substituting )N-[2-[N-phenyl-N-(2-methyl-1-propyl)amino]thiazol-4-ylcarbonyl]hydrazidefor(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-(2-hydrazinocarbonylthiazol-4-yl)-3′-methylbutyl]4-methylpentanamide,the title compound was prepared as a white solid (92 mg, 0.171 mmol). MS(ESI): 560.3 (M+Na)⁺.

Example 117

Preparation ofN-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 113(a)-113(f), except substitutinganiline for 4-aminobiphenyl in step (a), the title compound was preparedas a yellow solid (50 mg, 0.092 mmol). MS (ESI): 539.4 (M+H)⁺.

Example 118

Preparation ofN-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]-N′-[N-(3-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 112(a)-112(g), except substitutingN-(3-pyridinylmethoxycarbonyl)-L-leucine forN-(4-pyridinylmethoxycarbonyl)-L-leucine in step (g), the title compoundwas prepared as a white solid (93.8 mg, 53%). MS (ESI): 574.3 (M+H)⁺.

Example 119

Preparation ofN-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]-N′-[N-(2-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 112(a)-112(g), except substitutingN-(2-pyridinylmethoxycarbonyl)-L-leucine forN-(4-pyridinylmethoxycarbonyl)-L-leucine in step (g), the title compoundwas prepared as a white solid (149.7 mg, 85%). MS (ESI): 574.4 (M+H)⁺.

Example 120

Preparation ofN-(N-benzyloxycarbonyl-N-methyl-L-leucinyl)-N′-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 112(a)-112(g), except substitutingN-benzyloxycarbonyl-N-methyl-L-leucine forN-(4-pyridinylmethoxycarbonyl)-L-leucine in step (g), the title compoundwas prepared as a white solid (153.5 mg, 85%). MS (ESI): 609.3 (M+H)⁺.

Example 121

Preparation ofN-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]-N′-[N-(2-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 113(a)-113(f), except substitutinganiline for 4-aminobiphenyl in step (a) andN-(2-pyridinylmethoxycarbonyl)-L-leucine forN-(4-pyridinylmethoxycarbonyl)-L-leucine in step (f), the title compoundwas prepared as a white solid (40 mg). MS (ESI): 539.4 (M+H)⁺.

Example 122

Preparation ofN-[2-[N-(2-methylpropyl-N-phenylamino]thiazol-4-ylcarbonyl]-N′-[N-(3-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 113(a)-113(f), except substitutinganiline for 4-aminobiphenyl in step (a) andN-(3-pyridinylmethoxycarbonyl)-L-leucine forN-(4-pyridinylmethoxycarbonyl)-L-leucine in step (f), the title compoundwas prepared as a white solid (42 mg). MS (ESI): 539.4 (M+H)⁺.

Example 123

Preparation ofN-[2-(2-methoxyphenyl)thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

a) 2-trimethylstannylanisole

To a stirring solution of n-BuLi (2.6 mL, 2.5M in hexane, 6.42 mmol) indiethyl ether (2.5 mL) at −78C was added 2-bromoanisole (1.0 g, 5.35mmol) in diethyl ether (2 mL) dropwise. After stirring for 1 h at −78C,trimethyltin chloride (6.4 mL, 1.0M in THF, 6.42 mmol) was addeddropwise. The mixture was allowed to stir an additional 2 h while slowlywarming to room temperature. The mixture was then washed with saturatedaqueous NaHCO₃. The aqueous layer was extracted with diethyl ether (1×50mL) and the organic layers were combined, dried (MgSO₄), filtered andconcentrated. The residue was purified by column chromatography (silicagel, hexane) to yield the title compound as a colorless oil (1.11 g,76%). ¹HNMR (400 MHz, CDCl₃) δ 7.47 (d, 1H), 7.40 (t, 1H), 7.05 (t, 1H),6.90 (d, 1H), 3.36 (s, 3H), 0.34 (s, 9H).

b) ethyl 2-(2-methoxyphenyl)thiazole-4-carboxylate

A mixture of the compound of Example 112(b) (0.250 g, 1.06 mmol), thecompound of Example 83(a) (0.287 g, 1.06 mmol), andtetrakis(triphenylphosphine)palladium(0) (0.037 g, 0.0318 mmol) intoluene (2 mL) was stirred at reflux for 16 h. The mixture was dilutedwith ethyl acetate and washed with water and brine. The organic layerwas dried (MgSO₄), filtered and concentrated. The residue was purifiedby column chromatography (silica gel, ethyl acetate/hexane) to yield thetitle compound as a white solid (0.081 g, 29%). ¹HNMR (400 MHz, CDCl₃) δ8.54 (d, 1H), 8.22 (s, 1H), 7.45 (t, 1H), 7.11 (t, 1H), 7.05 (d, 1H),4.48 (q, 2H), 4.04 (s, 3H), 1.46 (t, 3H).

c)N-[2-(2-methoxyphenyl)thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 112(f)-112(g), except substitutingethyl 2-(2-methoxyphenyl)thiazole-4-carboxylate for ethyl2-(2-benzyloxyphenyl)thiazole-4-carboxylate in step (f), the titlecompound was prepared as a white solid. MS (ESI): 498.3 (M+H)⁺.

The above description fully discloses how to make and use the compoundsof the present invention. However, the present invention is not limitedto the particular embodiments described hereinabove, but includes allmodifications thereof within the scope of the following claims. Thevarious references to journals, patents and other publications which arecited herein comprise the state of the art and are incorporated hereinby reference as though fully set forth.

Example 124

Preparation of(2S,1′S)-N-[1′-(4-carboethoxythiazol-2-yl)-3′-methylbutyl]-4-methyl-2-(2-phenylbenzyloxycarbonyl)aminopentanamide

a)(2S,1′S)-2-(tert-butoxycarbonyl)amino-N-[1′-(4-carboethoxythiazol-2-yl)-3′-methylbutyl]-4-methylpentanamide

The compound of Example 8(c)(1.2 g, 3.5 mmol) was stirred at roomtemperature in neat TFA (2.96 g, 26.0 mmol) for 15 min. The solution wasthe concentrated in vacuo and redissolved in DMF (25 mL). To thestirring solution was added triethylamine (0.779 g, 7.7 mmol),BOC-Leu-OH (0.972 g, 3.9 mmol), 1-hydroxybenzotriazole (0.095 g, 0.7mmol), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(0.750 g, 3.9 mmol). After stirring at room temperature for 16 hours,the solution was diluted with ethyl actate and washed successively withwater (2×100 mL), NaHCO₃, and brine. The organic layer was dried(MgSO₄), filtered and concentrated. The residue was purified by columnchromatography (silica gel, ethyl acetatelhexane) to yield the titlecompound as a white solid (1.15 g, 72%). MS(ESI): 456.2 (M+H)⁺.

b)(2S,1′S)-N-[1′-(4-carboethoxythiazol-2-yl)-3′-methylbutyl]-4-methyl-2-(2-phenylbenzyloxycarbonyl)aminopentanamide

To a stirring solution of phosgene (1.5 mL, 2.9 mmol, 1.93M in toluene)at 0° C. was added 2-biphenylmethanol (0.486 g, 2.64 mmol) anddiisopropylethylamine (0.375 g, 2.9 mmol). The solution was allowed tostir at 0° C. for 30 min. In a separate reaction vessel, after stirringat room temperature for 10 min, the compound of Example 124(a) (0.150 g,0.330 mmol) dissolved in TFA (2.0 mL) was concentrated and redissolvedin DMF (3 mL). This solution was added to the 2-biphenylmethanolsolution followed by diisopropylethylamine (0.213 g, 1.65 mmol). Afterstirring at room temperature for 1 h, the solution was diluted withethyl acetate and washed successively with water and brine. The organiclayer was dried (MgSO₄), filtered and concentrated. The residue waspurified by column chromatography (silica gel, ethyl acetatethexane) toyield the title compound as a white solid (0.138 g, 74%). MS(ESI): 566.3(M+H)⁺.

Example 125

Preparation of(2S,1′S)-2-[(2-benzyl)benzyloxycarbonyl)]amino-N-[1′-(4-carboethoxythiazol-2-yl-)-3′-methylbutyl]-4-methylpentanamide

Following the procedure of Example 124(b), except substituting2-benzylbenzyl alcohol for 2-biphenylmethanol, the title compound wasprepared as a white solid (0.123 g, 64%). MS(ESI): 580.0 (M+H)⁺.

Example 126

Preparation of(2S,1′S)-N-[1′-(4-carboethoxythiazol-2-yl)-3′-methylbutyl]-4-methyl-2-(2-napthylmethoxycarbonyl)]aminopentanamide

Following the procedure of Example 124(b), except substituting2-naphthalenemethanol for 2-biphenylmethanol, the title compound wasprepared as a white solid (0.132 g, 74%). MS(ESI): 540.1 (M+H)⁺.

Example 127

Preparation of(2S,1′S)-N-[(4carboethoxythiazol-2)-3′-methylbutyl]-4-methyl-2-[(3-phenoxybenzyloxycarbonyl)]aminopentanamide

Following the procedure of Example 124(b), except substituting3-phenoxybenzyl alcohol for biphenylmethanol, the title compound wasprepared as a white solid (0.107 g, 56%). MS(ESI): 581.9 (M+H)⁺.

Example 128

Preparation of(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-[2-(2-benzylguanidinyl)thiazol-4-yl]-3′-methylbutyl]-4-methylpentanamide

a) S-methyl Dithiobiuret Hydroiodide Salt

To a stirring solution of dithiobiuret (5.0 g, 37 mmol) in THF (75 mL)was added iodomethane (13.1 g, 92.5 mmol, 5.76 μl). After stirring atroom temperature for 22 h, the solution was diluted with 150 mL oftoluene and allowed to stand at 0° C. for 3 h. The crystals werecollected by filtration and washed with cold 2:1 toluene/THF, then driedin vacuo to give the title compound as a white solid (8.7 g, 85%).MS(ESI): 149.9 (M+H)⁺.

b) 3-benzylguanidinyl Thiourea

The compound of Example 128(b) (4.35 g, 15.7 mmol) was dissolved inisopropanol (80 mL) and benzylamine (1.77 g, 16.5 mmol, 1.8 mL) wasadded and the mixture was heated at reflux for 16 h. The hot solutionwas filtered and the filtrate was cooled to 0° C. After 5 h, the solidwas collected by filtration and washed twice with cold iospropanol, thendried in vacuo to provide the title compound as a white solid (2.59 g,61%). MS(ESI): 209.2 (M+H)⁺.

c)(2S,1′S)-2-(benzyloxycarbonyl)amino-N-[1′-[2-(2-benzylguanidinyl)thiazol-4-yl]-3′-methylbutyl]4-methylpentanamide

Following the procedure of Example (b), except substituting3-benzylguanidinyl thiourea for ethyl thiooxamate, the title compoundwas prepared as a white solid (102 mg, 79%). MS(ESI): 565.1 (M+H)⁺.

Example 129

Preparation of(1S)-N-[4-[1-(N-benzyloxycarbonylamino)-3-methylbutyl]thiazol-2-ylcarbonyl]-N-methyl-N′-(N-benzyloxycarbonyl-L-leucinyl)hydrazide

Following the procedure of Example 26(a)-26(d), except substitutingmethyl hydrazine for hydrazine in step (c), the title compound wasprepared as a white solid. MS(ESI): 624.1 (M+H)⁺.

Example 130

Preparation of(1S)-N-[4-1-(N-benzyloxycarbonylamino)-3-methylbutyl]thiazol-2-ylcarbonyl]-N′-(N-benzyloxycarbonyl-L-leucinyl)-N′-methylhydrazide

a) N-(N-benzyloxycarbonyl-L-leucinyl)-N-methylhydrazide

Following the procedure of Example 26(c), except substitutingN-benzyloxycarbonyl-L-leucine methyl ester for(1S)-1-benzyloxycarbonylamino-1-(2-carboethoxythiazol-4-yl)-3-methylbutaneand methyl hydrazine for hydrazine, the title compound was prepared.

b)(1S)-1-benzyloxycarbonylamino-1-(2-carboxythiazol-4-yl)-3-methylbutane

The compound of Example 26(c)(0.57 g., 1.5 mmol) was dissolved intetrahydrofuran and treated with an excess of 1.0N sodium hydroxide. Themixture was allowed to stir for 4 hours, and was quenched with 1.0Ncitric acid. The solvent was evaporated and the aqueous layer extractedthree times with dichloromethane. The organic layers were combined andevaporated to give the acid as a white foam (0.55 g, 100%).

c)(1S)-N-[4-[1-(N-benzyloxycarbonylamino)-3-methylbutyl]thiazol-2-ylcarbonyl]-N′-(N-benzyloxycarbonyl-L-leucinyl)-N′-methylhydrazide

Following the procedure of Example 28(e), except substitutingN-(N-benzyloxycarbonyl-L-leucinyl)-N-methylhydrazide for(1S)-1-(benzyloxycarbonyl)amino-1-(4-carboethoxythiazol-2-yl)-3-methylbutaneand(1S)-1-benzyloxycarbonylamino-1-(2-carboxythiazol-4-yl)-3-methylbutanefor N-benzyloxycarbonyl-L-leucine, the title compound was prepared as awhite solid. MS (ESI): 624.2 (M+H)⁺.

Example 131

Preparation ofN-(N-benzyloxycarbonyl-L-leucinyl)-N′-(N-benzyloxycarbonyl-L-leucinyl)-L-alanylhydrazide

Following the procedure of Example 27(a)-27(c), except substitutingL-alanine methyl ester for L-leucine methyl ester in step (a), the titlecompound was prepared as a white solid (225 mg, 42%). MS(ESI): 598.1(M+H)⁺.

Example 132

Preparation ofN-(N-benzyloxycarbonyl-L-leucinyl)-N′-(N-benzyloxycarbonyl-L-leucinyl)glycinylhydrazide

Following the procedure of Example 27(a)-27(c), except substitutingglycine methyl ester for L-leucine methyl ester in step (a), the titlecompound was prepared as a white solid (307 mg, 42%). MS(ESI): 584.1(M+H)⁺.

Example 133

Preparation of(1S)-N-[2-1-(N-benzyloxycarbonylamino)-3-methylbutyl]-1,3,4-triazol-5-ylcarbonyl]-N′-(N-benzyloxycarbonyl-L-leucinyl)hydrazide

a) Ethyl Oxalamidrazonate

To a solution of ethyl thiooxamate (3.0 g, 22.6 mmol) in ethanol (50 mL)was added hydrazine hydrate (1.13 g, 22.6 mmol, 1.09 mL). The mixturewas allowed to stir for 3 hours at room temperature, while ventingthrough a scrubber of concentrated sodium hydroxide solution. Thesolution was allowed to stand for 16 hours and the ethanol wasevaporated. The residue was boiled in 30% dichloromethane in petroleumether, filtered, and recrystallized to give the desired compound as atan solid. (0.264 g, 9%).

b) (1S)-1-benzyloxycarbonylamino--(2-carboethoxy-1,3,4-triazol-5-yl)-3-methylbutane

N-benzyloxycarbonyl-L-leucine (0.535 g, 2.0 mmol) was stirred in THF at−5° C. Ethyl chloroformate (0.23 mL, 2.4 mmol) and triethylamine (0.25g, 2.4 mmol, 0.34 mL) were added. The compound of Example 10(a) (0.264g, 2.0 mmol) was then added and the mixture was allowed to stir at roomtemperature overnight. The solvents were evaporated and the residue wasdissolved in xylenes and heated to 200° C. using a Dean-Stark apparatus.The heating was stopped after 4 hours and the solution was evaporated toa residue which was chromatographed (silica gel, 40% ethyl acetate inhexane) to give the title compound as a white solid (0.498 g, 69%).¹HNMR(400 MHz, CDCl₃) δ 7.20 (m, 5H), 5.71 (d, 1H), 5.04 (s, 2H), 4.99(dd, 1H), 4.36 (q, 2H), 1.8 (m, 2H), 1.59 (m, 1H), 1.31 (t, 3H),0.83(dd, 6H).

c)(1S)-1-benzyloxycarbonylamino-1-(2-hydrazinocarbonyl-1,3,4-triazol-5-yl)-3-methylbutane

Following the procedure of Example 26(c)-26(d), except substituting(1S)-1-benzyloxycarbonylamino-1-(2-carboethoxy-1,3,4triazol-5-yl)-3-methylbutanefor(1S)-1-benzyloxycarbonylamino-1-(2-carboethoxythiazol-4-yl)-3-methylbutanein step (c), the tide compound was prepared. MS (ESI): 594.5 (M+H)⁺.

Example 134

Preparation of(1S)—N-(N-acetyl-L-leucinyl)-N′-[2-[1-(N-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 28(a)-28(e), except substitutingN-acetyl-L-leucine for N-benzyloxycarbonyl-L-leucine in step (e), thetitle compound was prepared as a white solid (95 mg, 67%). MS(ESI):518.0 (M+H)⁺.

Example 135

Preparation of(1S)-N-(N-benzyloxycarbonyl-L-alanyl)-N′-[2-[1-(N-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 28(a)-28(e), except substitutingN-benzyloxycarbonyl-L-alanine for N-benzyloxycarbonyl-L-leucine in step(e), the title compound was prepared as a white solid (129 mg, 82%).MS(ESI): 568.1 (M+H)⁺.

Example 136

Preparation of(1S)-N-(N-acetyl-L-alanyl)-N′-[2-[1-(N-benzyloxycarbonylamino-3-methylbutyl]thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 28(a)-28(e), except substitutingN-acetyl-L-alanine for N-benzyloxycarbonyl-Leucine in step (e), thetitle compound was prepared as a white solid (74 mg, 57%). MS(ESI):498.1 (M+Na)⁺.

Example 137

Preparation of(1S)-N-(N-acetyl)-N′-[2-[1-(N-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 28(a)-28(e), except substitutingacetic acid for N-benzyloxycarbonyl-L-leucine in step (e), the titlecompound was prepared as a white solid (87 mg, 78%). MS(ESI): 405.1(M+H)⁺.

Example 138

Preparation of(1S)-N-[2-[1-(N-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 28(a)-28(e), except substitutingN-(4-pyridinylmethoxycarbonyl)-L-leucine forN-benzyloxycarbonyl-L-leucine in step (e), the title compound wasprepared as a white solid (121 mg, 72%). MS(ESI): 611.0 (M+H)⁺.

Example 139

Preparation of(1S)-N-[2-[1-(N-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[N-(2-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 28(a)-28(e), except substitutingN-(2-pyridinylmethoxycarbonyl)-L-leucine forN-benzyloxycarbonyl-L-leucine in step (e), the title compound wasprepared as a white solid (125 mg, 65%). MS (ESI): 611.2 (M+H)⁺.

Example 140

Preparation of(1S)-N-[2-[1-(N-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-(N-benzyloxycarbonyl-N-methyl-L-leucinyl)hydrazide

Following the procedure of Example 28(a)-28(e), except substitutingN-benzyloxycarbonyl-N-methyl-L-leucine for N-benzyloxycarbonyl-L-leucinein step (e), the title compound was prepared as a white solid (78 mg,50%). MS (ESI): 624.3 (M+H)⁺.

Example 141

Preparation of(1S)-N-[2-[1-(N-benzyloxycarbonyl-N-methylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 28(a)-28(e), except substitutingN-benzyloxycarbonyl-N-methyl-L-leucine forN-tert-butoxycarbonyl-L-leucine in step (a) andN-(2-pyridinylmethoxycarbonyl)-L-leucine forN-benzyloxycarbonyl-L-leucine in step (e), the title compound wasprepared as a white solid (120 mg, 72%). MS (ESI): 625.3 (M+H)⁺.

Example 142

Preparation of(1′S)-N-(N-benzyloxycarbonyl-L-leucinyl)-N′-[2-[1-(N-benzyloxycarbonyl-N-methylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 28(a)-28(e), except substitutingN-benzyloxycarbonyl-N-methyl-L-leucine forN-tert-butoxycarbonyl-L-leucine in step (a), the title compound wasprepared as a white solid (95 mg, 74%). MS (ESI): 624.3 (M+H)⁺.

Example 143

Preparation of(1S)-N-[2-[1-(N-benzyloxycarbonyl-N-methylamino)-3-methylbutyl]thiazol-4-yl]carbonyl-N′-(N-benzyloxycarbonyl-N-methyl-L-leucinyl)hydrazide

Following the procedure of Example 28(a)-28(e), except substitutingN-benzyloxycarbonyl-N-methyl-L-leucine forN-tert-butoxycarbonyl-L-leucine in step (a) andN-benzyloxycarbonyl-N-methyl-L-leucine for N-benzyloxycarbonyl-L-leucinein step (e), the title compound was prepared as a white solid (129 mg,59%). MS (ESI): 683.3 (M+H)⁺.

Example 144

Preparation of(1S)-N-[2-1-(N-methylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

a)(1S)-N-[2-[1-(N-tert-butoxycarbonyl-N-methylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 28(a)-28(e), except substitutingN-tert-butoxycarbonyl-N-methyl-L-leucine forN-tert-butoxycarbonyl-L-leucine in step (a) andN-(4-pyridinylmethoxycarbonyl-L-leucine forN-benzyloxycarbonyl-L-leucine in step (e), the title compound wasprepared as a white solid. MS (ESI): 591.4 (M+H)⁺.

b)(1S)-N-[2-[1-(N-methylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

To a solution of the compound of Example 21(a) in methylene chloride (10mL) was added trifluoroacetic acid (3 mL). After stirring one hour atroom remperature the solution was concentrated and the residue wasredissolved in methylene chloride, washed with saturated aqueous sodiumbicarbonate, dried over MgSO₄ and concentrated to afford the titlecompound as a white solid (259 mg, 68% for two steps). MS (ESI): 491.4(M+H)⁺.

Example 145

Preparation of(1S)-N-[2-[1-(N-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[N-(tert-butoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 28(a)-28(e), except substitutingN-tert-butoxycarbonyl-L-leucine for N-benzyloxycarbonyl-L-leucine instep (e), the title compound was prepared as a white solid (293 mg,74%). MS (ESI): 576.4 (M+H)⁺.

Example 146

Preparation of(1S)-N-[2-[1-(benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[N-(tert-butoxycarbonyl)-N-methyl-L-leucinyl]hydrazide

Following the procedure of Example 28(a)-28(e), except substitutingN-tert-butoxycarbonyl-N-methyl-L-leucine forN-benzyloxycarbonyl-L-leucine in step (e), the title compound wasprepared as a white solid (120 mg, 87%). MS (ESI): 590.3 (M+H)⁺.

Example 147

Preparation of(1S)-N-[2-[1-(N-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-(N-methyl-L-leucinyl)hydrazide

Following the procedure of Example 144(b), except substituting(1S)-N-[2-[1-(N-benzyloxycarbonylamino)₃-methylbutyl]thiazol-4-ylcarbonyl]-N′-[N-(tert-butoxycarbonyl)-N-methyl-L-leucinyl]hydrazidefor(1S)-N-[2-[1-(N-tert-butoxycarbonyl-N-methylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide,the title compound was prepared as a white solid (40 mg, 80%). MS (ESI):490.3 (M+H)⁺.

Example 148

Preparation of(1S)-N-[2-[1-(N-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-(L-leucinyl)hydrazide

Following the procedure of Example 144(b), except substituting(1S)-N-[2-[1-(N-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[N-(tert-butoxycarbonyl)-L-leucinyl]hydrazidefor(1S)-N-[2-[1-(N-tert-butoxycarbonyl-N-methylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)L-leucinyl]hydrazide,the title compound was prepared as a white solid (39 mg, 100%). MS(ESI): 476.4 (M+H)⁺.

Example 149

Preparation of(1S)-N-[2-[1-(N-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[N-(4-imidazolylacetyl)-L-leucinyl]hydrazide

Following the procedure of Example 28(e), except substituting(1S)-N-[2-[1-(N-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-(L-leucinyl)hydrazidefor(1S)-1-(benzyloxycarbonyl)amino-1-(4-carboethoxythiazol-2-yl)-3-methylbutaneand 4-imidazoleacetic acid for N-benzyloxycarbonyl-L-leucine, the titlecompound was prepared as a white solid (50 mg, 47%). MS (ESI): 584.4(M+H)⁺.

Example 150

Preparation of(1S)-N-[2-[1-N-benzyloxycarbonyl-N-methylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[N-(3-pyridinylmethoxycarbonyl)-N-methyl-L-leucinyl]hydrazide

a) N-methyl-L-leucine Methyl Ester

N-methyl-L-leucine (1.3 g, 8.95 mmol) was dissolved in 4M HCl,1,4-dioxane (10 mL) and methanol (10 mL). The solution was stirredovernight at room temperature, then concentrated to afford the titlecompound as a white solid (100%). MS (ESI): 160.0 (M+H)⁺.

b) N-methyl-N-(3-pyridinylmethoxycarbonyl)-L-leucine Methyl Ester

To a stirring solution of phosgene in toluene (5.63 mL, 6.025 mmol) inmethylene chloride (10 mL), cooled to 0° C., was added dropwise asolution of N-methyl-L-leucine methyl ester (673 mg, 4.63 mmol) andpyridine (1.10 g, 0.97 mL, 13.89 mmol) in methylene chloride (4 mL). Thesolution was stirred at 0° C. for 2 hours. A solution of 3-pyridylcarbinol (0.56 g, 5.09 mmol, 0.49 mL) was then added and the reactionmixture was stirred at room temperature for 5 hours. The solution wasconcentrated, redissolved in ethyl acetate, washed with water, dried(MgSO₄), filtered and concentrated. The crude residue was purified bycolumn chromatography on silica gel (6% methanol in methylene chloride)to afford the title compound as a yellow oil (88 mg, 7%). MS (ESI):295.4 (M+H)⁺.

c) N-methyl-N-(3-pyridinylmethoxycarbonyl)-L-leucine

Following the procedure of Example 130(b), except substitutingN-methyl-N-(3-pyridinylmethoxycarbonyl)-L-leucine methyl ester(1S)-1-benzyloxycarbonylamino-1-(2-hydrazinocarbonylthiazol-4-yl)-3-methylbutane,the title compound was prepared as an orange solid (84 mg, 100%). MS(ESI): 281.3 (M+H)⁺.

d)(1S)-N-[2-[1-(N-benzyloxycarbonyl-N-methylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[N-(3-pyridinylmethoxycarbonyl)-N-methyl-L-leucinyl]hydrazide

Following the procedure of Example 28(a)-28(e), except substitutingN-benzyloxycarbonyl-N-methyl-L-leucine forN-tert-butoxycarbonyl-L-leucine in step (a) andN-methyl-N-(3-pyridinylmethoxycarbonyl)-L-leucine forN-benzyloxycarbonyl-L-leucine in step (e), the title compound wasprepared as a white solid (55 mg, 38%). MS (ESI): 639.4 (M+H)⁺.

Example 151

Preparation of(1S)-N-[2-[1-(N-benzyloxycarbonyl-N-methylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[N-(3-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 28(a)-28(e), except substitutingN-benzyloxycarbonyl-N-methyl-L-leucine forN-tert-butoxycarbonyl-L-leucine in step (a) andN-(3-pyridinylmethoxycarbonyl)-L-leucine forN-benzyloxycarbonyl-L-leucine in step (e), the title compound wasprepared as a white solid (31 mg, 34%). MS (ESI): 625.4 (M+H)⁺.

Example 152

Preparation of(1S)—N-[2-[1-(N-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[N-(3-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 28(a)-28(e), except substitutingN-(3-pyridinylmethoxycarbonyl)-L-leucine forN-benzyloxycarbonyl-L-leucine in step (e), the title compound wasprepared as a white solid (63 mg, 42%). MS (ESI): 611.5 (M+H)⁺.

Example 153

Preparation of(1′S)-N-(N-benzyloxycarbonyl-L-leucinyl)-N′-(1-benzyloxycarbonyl)-N′-[2-[1-(N-methylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-methylhydrazide

Following the procedure of Example 28(a)-28(e), except substitutingN-benzyloxycarbonyl-N-methyl-L-leucine forN-tert-butoxycarbonyl-L-leucine in step (a) and methyl hydrazine forhydrazine in step (e), the title compound was prepared as a white solid(80 mg, 70%). MS (ESI): 660.4 (M+Na)⁺.

Example 154

Preparation of(1S)-N-[2-[1-(N-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[N-(2-pyridinylmethoxycarbonyl)-N-methyl-L-leucinyl]hydrazide

a) N-methyl-N-(2-pyridinylmethoxycarbonyl)-L-leucine Methyl Ester

N-(2-pyridinylmethoxycarbonyl)-L-leucine methyl ester (490 mg, 1.75mmol) was dissolved in THF (7.0 mL) and methyl iodide (0.435 mL, 6.99mmol) was added. The reaction mixture was cooled to 0° C. in a flaskprotected from moisture. Sodium hydride dispersion (236 mg, 2.62 mmol)was added cautiously and the suspension was stirred for 5 hours at roomtemperature. Ethyl acetate was then added, followed by water, dropwise.The solution was concentrated in vacuo, and the oily residue partitionedbetween ether and water. The organic layer was washed with saturatedaqueous NaHCO₃ and the combined aqueous extracts acidified to pH 3 withcitric acid. The product was extracted with ethyl acetate, the extractwas washed with water, 5% aqueous sodium thiosulfate and water, dried(MgSO₄), filtered and concentrated. The crude product was purified bycolumn chromatography on silca gel (ethyl acetate/hexane, 3:1) to give ayellow oil (235 mg, 46%). MS (ESI): 295.4 (M+H)⁺.

b) N-methyl-N-(2-pyridinylmethoxycarbonyl)-L-leucine

Following the procedure of Example 130(b), except substitutingN-methyl-N-(2-pyridinylmethoxycarbonyl)-L-leucine methyl ester(1S)-1-benzyloxycarbonylamino-1-(2-hydrazinocarbonylthiazol-4-yl)-3-methylbutane,the title compound was prepared as a white solid (223 mg, 100%). MS(ESI): 281.3 (M+H)⁺.

c)(1S)-N-[2-[1-(N-benzyloxycarbonylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[N-(2-pyridinylmethoxycarbonyl)-N-methyl-L-leucinyl]hydrazide

Following the procedure of Example 28(a)-28(e), except substitutingN-benzyloxycarbonyl-N-methyl-L-leucine forN-tert-butoxycarbonyl-L-leucine in step (a) andN-methyl-N-(2-pyridinylmethoxycarbonyl)-L-leucine forN-benzyloxycarbonyl-L-leucine in step (e), the title compound wasprepared as a white solid (50 mg, 44%). MS (ESI): 639.5 (M+H)⁺.

Example 155

Preparation of(1S)-N-[2-[1-(N-benzyloxycarbonyl-N-methylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-N-methyl-L-leucinyl]hydrazide

a) L-leucine Tert-Butyl Ester Isocyanate

L-leucine tert-butyl ester hydrochloride (10.185 g, 45.5 mmol) wasdissolved in methylene chloride (100 mL), cooled to 0° C. and pyridine(12.7 mL, 182.0 mmol) was added, then phosgene in benzene (47 mL, 59.1mmol). The solution was stirred at 0° C. for 2 hours. The reactionmixture was washed two times with 300 mL of cold 0.5 M aqueous HCl. Eachaqueous layer was exctracted with 100 mL methylene chloride. Thecombined organic phases were washed with a mixture of saturated aqueousNaCl solution and crushed ice, dried over MgSO₄, filtered andconcentrated to afford the isocyanate as a yellow liquid (5.37 g, 55%).

b) N-(4-pyridinylmethoxycarbonyl)-L-leucine Tert-Butyl Ester

The compound of Example 155(a) (3.05 g, 14.32 mmol) and 4-pyridylcarbinol (1.56 g, 14.32 mmol) were dissolved in toluene (80 mL) andheated at reflux overnight. The solution was concentrated in vacuo andthe residue was purified by column chromatography on silica gel (ethylacetate/hexane, 3:1) to afford the title compound as a colorless oil(2.945 g, 64%). MS (ESI): 323.4 (M+H)⁺.

c) N-methyl-N-(4-pyridinylmethoxycarbonyl)-L-leucine Tert-Butyl Ester

Following the procedure of Example 154(a), except substitutingN-(4-pyridinylmethoxycarbonyl)-L-leucine tert-butyl ester forN-(2-pyridinylmethoxycarbonyl)-L-leucine methyl ester, the titlecompound was prepared as a yellow liquid (2.038 g, 68% yield). MS (ESI):337.5 (M+H)⁺.

d) N-methyl-N-(4pyridinylmethoxycarbonyl)-L-leucine

Following the procedure of Example 144(b), except substitutingN-methyl-N-(4-pyridinylmethoxycarbonyl)-L-leucine tert-butyl ester for(1S)-N-[2-[1-(N-tert-butoxycarbonyl-N-methylamino)-3-methylbutyl]thiazolylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide,the title compound was prepared as a white solid (343 mg, 72% yield). MS(ESI): 281.3 (M+H)⁺.

e)(1S)-N-[2-[1-(N-benzyloxycarbonyl-N-methylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[N-(4pyridinylmethoxycarbonyl)-N-methyl-L-leucinyl]hydrazide

Following the procedure of Example 28(a)-28(e), except substitutingN-benzyloxycarbonyl-N-methyl-L-leucine forN-tert-butoxycarbonyl-L-leucine in step (a) andN-methyl-N-(4-pyridinylmethoxycarbonyl)-L-leucine forN-benzyloxycarbonyl-L-leucine in step (e), the title compound wasprepared as a white solid (50 mg, 44%). MS (ESI): 639.5 (M+H)⁺.

Example 156

Preparation of 2,2′-[N,N′-[bis-(N-acetyl-L-leuccinyl)]]carbohydrazide

Following the procedure of Example 29, except substitutingN-acetyl-L-leucine for N-benzyloxycarbonyl-L-leucine, the title compoundwas prepared as a pale yellow solid (0.153 g, 23%). MS(ESI): 401.3(M+H)⁺.

Example 157

Preparation of2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-(4-methylpentanoyl)]carbohydrazide

a) N-benzyloxycarbonyl-L-leucine Methyl Ester

To a stirring solution of L-leucine methyl ester (2.0 g, 11.0 mmol) in1,4-dioxane (20 mL) was added aqueous Na₂CO₃ solution (12.1 mL, 2M inH₂O) followed by benzylchloroformate (1.96 g, 11.5 mmol). The mixturestirred at room temperature for 4 h then partitioned between ethylacetate and water. The organic layer was washed with saturated brine,dried (MgSO₄), filtered and concentrated to yield the title compound asa colorless oil (3.1 g, 100%). ¹HNMR (400 MHz, CDCl₃) δ 7.34 (m, 5H),5.27 (d, 1H), 5.12 (s, 2H), 4.41 (s, 2H), 3.75 (s, 3H), 1.65 (m, 3H),0.96 (m, 6H).

b) N-(N-benzyloxycarbonyl-L-leucinyl)hydrazide

To a stirring solution of the compound of Example 157(a) (3.1 g, 11.0mmol) in methanol (15 mL) was added hydrazide hydrate (5.9 g, 118 mmol,5.7 mL). The solution stirred at room temperature for 16 h thenconcentrated to yield the title compound as an off-white solid (3.1 g,100%). MS(ESI): 280.2 (M+H)⁺.

c) 1-benzyloxycarbonylamino-3-methyl-1-(1,3,4-oxadiazol-2-on-5-yl)butane

To a stirring solution of the compound of Example 157(b) (3.0 g, 10.8mmol) in toluene (50 mL) was added phosgene (56 mL, 1.93M in toluene).The solution was heated at reflux for 4 h, then concentrated to yieldthe title compound as a pale yellow foam (3.15 g, 96%). MS(ESI): 306.1(M+H)⁺.

d) N-(4-methylpentanoyl)hydrazide

To a stirring solution of ethyl isocaproate (2.0 g, 13.8 mmol) inethanol (25 mL) was added hydrazine monohydrate (6.9 g, 138 mmol, 6.7mL). After stirring at room temperature for 48 h, the solution wasconcentrated to yield the title compound as a white solid. (1.8 g,100%). ¹HNMR (400 MHz, CDCl₃) δ 7.48 (s br, 1H), 3.62 (s br, 2H), 2.13(t, 2H), 1.51 (m, 3H), 0.85 (d, 6H).

e)2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-(4-methylpentanoyl)]carbohydrazide

The compounds of Example 157(c) (0.100 g, 0.325 mmol) and Example 34(d)(0.042 g, 0.325 mmol) were combined and dissolved in ethanol (1 mL). Thesolution was heated at reflux for 24 hours, then concentrated to a solidyellow residue which was washed with cool methylene chloride to yieldthe title compound as a white solid (0.053 g, 37%). MS(ESI): 436.2(M+H)⁺.

Example 158

Preparation of2,2′-[N,N′-[bis-(N-benzyloxycarbonyl-N-methyl-L-leucinyl)]]carbohydrazide

Following the procedure of Example 29, except substitutingN-benzyloxycarbonyl-N-methyl-L-leucine for substitutingN-benzyloxycarbonyl-L-leucine, the title compound was prepared withpurification by column chromatography (silica gel,methanol/dichloromethane) as a white foam (0.236 g, 23%). MS (MH⁺):613.2.

Example 159

Preparation of2-[N-(N-acetyl-L-leucinyl)]-2′-[N′-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide)

a) 2-[N-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide

To a stirring solution of the compound of Example 157(c) (3.15 g, 10.3mmol) in methanol (2 mL) was added hydrazine hydrate (5.0 g, 100 mmol,4.8 mL). After stirring at room temperature for 24 h, the solution wasconcentrated to yield the title compound as a pale yellow foam (3.471 g,100%). MS(ESI): 338.2 (M+H)⁺.

b)2-[N-(N-acetyl-L-leucinyl)]-2′-[N′-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide

To a stirring solution of the compound of Example 159(a) (0.100 g, 0.297mmol), N-acetyl-L-leucine (0.054 g, 0.312 mmol) and1-hydroxybenzotriazole (0.008 g, 0.0594 mmol) in DMF (2 mL) was added1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.060 g,0.312 mmol). After stirring at room temperature for 16 h, the solutionwas poured into water and extracted with ethyl acetate. The organiclayer was dried (MgSO₄), filtered and concentrated. The residue waspurified by column chromatography (silica gel, methanol/dichloromethane)to yield the title compound as a white solid (0.052 g, 36%). MS(ESI):493.1 (M+H)⁺.

Example 160

Preparation of2,2′-[N,N′-[bis-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl)]]]carbohydrazide

Following the procedure of Example 29, except substitutingN-(4-pyridinylmethoxycarbonyl)-L-leucine forN-benzyloxycarbonyl-L-leucine, the title compound was prepared as awhite solid (199 mg, 64%). MS(ESI): 587.1 (M+H)⁺.

Example 161

Preparation of2,2′-[N,N′-[bis-[N-(2-pyridinylmethoxycarbonyl)-L-leucinyl)]]]carbohydrazide

Following the procedure of Example 29, except substitutingN-(2-pyridinylmethoxycarbonyl)-L-leucine forN-benzyloxycarbonyl-L-leucine, the title compound was prepared as awhite solid (263 mg, 81%). MS(ESI): 587.1 (M+H)⁺.

Example 162

Preparation of2[-N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-[N-(2-pyridinylmethoxycarbonyl)-L-leucinyl]]carbohydrazide

Following the procedure of Example 159(a)-159(b) except substitutingN-(2-pyridinylmethoxycarbonyl)-L-leucine lithium salt forN-acetyl-L-leucine in step (b), the title compound was prepared as awhite solid (0.040 g, 15%). MS(ESI): 586.3 (M+H)⁺.

Example 163

Preparation of2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl)]]carbohydrazide

Following the procedure of Example 159(a)-159(b) except substitutingN-(4-pyridinylmethoxycarbonyl)-L-leucine lithium salt forN-acetyl-L-leucine in step (b), the title compound was prepared as awhite solid (0.045 g, 17%). MS(ESI)-586.3 (M+H)⁺.

Example 164

Preparation of2-[N-(N-benzyloxycarbonyl-L-leucinyl)-2′-[N′-[N-(3-pyridinylmethoxycarbonyl)-L-leucinyl)]]carbohydrazide

Following the procedure of Example 159(a)-159(b) except substitutingN-(3-pyridinylmethoxycarbonyl)-L-leucine lithium salt forN-acetyl-L-leucine in step (b), the title compound was prepared as awhite solid (0.084 g, 32%). MS(ESI): 586.3 (M+H)⁺.

Example 165

Preparation of2,2′-[N,N′-[bis-(N-benzyloxycarbonyl-L-leucinyl)]]-2-(N-methyl)carbohydrazide

a) N-methyl-N-(N-benzyloxycarbonyl-L-leucinyl)hydrazide

To a stirring solution of N-benzyloxycarbonyl-L-leucine methyl ester(2.2 g, 8.15 mmol) in methanol (4 mL) was added methylhydrazine (3.7 g,80 mmol). After stirring at room temperature for 16 h, the solution wasconcentrated to yield the title compound as a yellow solid (2.14 g, 7.3mmol). MS(ESI): 294.1 (M+H)⁺.

b)2,2′-[N,N′-[bis-(N-benzyloxycarbonyl-L-leucinyl)]]-2-(N-methyl)carbohydrazide

The compound of Example 157(c) (0.250 g, 0.819 mmol) and the compound ofExample 165(a) (0.240 g, 0.819 mmol) were combined, dissolved in ethanoland heated at reflux for 24 h. The solution was concentrated and theresidue purified by column chromatography (silica gel,methanol/dichloromethane) to yield the title compound as a white solid(0.060 g, 12%). MS(ESI): 599.1 (M+H)⁺.

Example 166

Preparation of2,2′-[N,N′-[bis-[N-(3-pyridinylmethoxycarbonyl)-L-leucinyl]]]carbohydrazide

Following the procedure of Example 29, except substitutingN-(3-pyridinylmethoxycarbonyl)-L-leucine forN-benzyloxycarbonyl-L-leucine, the title compound was prepared as awhite solid (157 mg, 48%). MS(ESI): 587.0 (M+H)⁺.

Example 167

Preparation of1-N-benzyl)-2,2′-[N,N′-[bis-(N-benzyloxycarbonyl-L-leucinyl)]]carbohydrazide

a) N-benzylidene-N′-(N-benzyloxycarbonyl-L-leucinyl)hydrazide

To a solution of the compound of Example 157(b) (1 g, 3.5 mmol) inethanol (30 mL) was added benzaldehyde (0.33 mL, 3.2 mmol). Theresulting mixture was heated at reflux for 4 h. The mixture wasconcentrated in vacuo then purified by flash chromatography (silica gel,10-50% EtOAc/hexane) to yield the title compound as a solid (0.31 g,23%). MS(ESI): 368.0 (M+H)⁺.

b) N-benzyl-N′-(N-benzyloxycarbonyl-L-leucinyl)hydrazide

To a cooled solution of compound of Example 167(a) (0.24 g, 0.65 mmol)in THF (5 mL) was added borane tetrahydrofuran complex (0.65 mL, 0.65mmol; 1M solution in THF). The resulting mixture was stirred at roomtemperature for 4 h then concentrated in vacuo and diluted with ethylacetate, washed with water, saturated brine, dried (MgSO₄), filtered andconcentrated in vacuo to give the title compound as a white solid (0.25g, 89%). MS(ESI): 370.0 (M+H)⁺.

c)1-benzyloxycarbonylamino-3-methyl-1-(3-benzyl-1,3,4-oxadiazol-2-on-5-yl)butane

Following the procedure of Example 157(c), except substitutingN-benzyl-N′-(N-benzyloxycarbonyl-L-leucinyl)hydrazide forN-(N-benzyloxycarbonyl-L-leucinyl)hydrazide, the title compound wasprepared as an oil (0.02 g, 83%). MS(ESI): 396.0 (M+H)⁺.

d) 1-(N-benzyl)-2-[N-(N-benzyloxycarbonyl-L-leucinyl)]carbohydrazide

Following the procedure of Example 159(a), except substituting1-benzyloxycarbonylamino-3-methyl-1-(3-benzyl-1,3,4-oxadiazol-2-on-5-yl)butanefor1-benzyloxycarbonylamino-3-methyl-1-(1,3,4-oxadiazol-2-on-5-yl)butane instep (a), the title compound was prepared as a solid (0.013 g, 62%).MS(ESI): 428.0 (M+H)⁺.

d)1-(N-benzyl)-2,2′-N,N′-[bis-(N-benzyloxycarbonyl-L-leucinyl)]]carbohydrazide

Following the procedure of Example 159(a)-159(b), except substituting1-benzyloxycarbonylamino-3-methyl-1-(3-benzyl-1,3,4-oxadiazol-2-on-5-yl)butanefor1-benzyloxycarbonylamino-3-methyl-1-(1,3,4-oxadiazol-2-on-5-yl)butane instep (a), the tide compound was prepared as white solid (13 mg, 86%).MS(ESI): 675.0 (M+H)⁺.

Example 168

Preparation of2-[N-(N-benzyloxycarbonyl-L-leucinyl)]-2′-[N′-(N-benzyloxycarbonyl-N-methyl-L-leucinyl)]carbohydrazide

Following the procedure of Example 159(a)-159(b) except substitutingN-benzyloxycarbonyl-N-methyl-L-leucine for N-acetyl-L-leucine in step(b), the title compound was prepared as a white solid (0.141 g, 53%).MS(ESI): 599.4 (M+H)⁺.

Example 169

Preparation ofN-[2-(1-naphthyl)thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 112(a)-112(g), except substituting1-naphthyl boronic acid for 2-benzyloxyphenyl boronic acid in step (e),the title compound was prepared as a white solid (0.094 g, 58%).MS(ESI): 518.4 (M+H)⁺.

Example 170

Preparation ofN-[2-(2-biphenyl)thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 112(a)-112(g), except substituting2-biphenylboronic acid for 2-benzyloxyphenyl boronic acid in step (e),the title compound was prepared as a white solid (0. 100 g, 43%).MS(ESI): 544.3 (M+H)⁺.

Example 171

Preparation ofN-(N-benzyloxycarbonyl-N-methyl-L-leucinyl)-N′-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 116(a)-116(b), except substitutingN-benzyloxycarbonyl-N-methyl-L-leucine forN-(4-pyridinylmethoxycarbonyl)-L-leucine in step (b), the title compoundwas prepared as a white solid (40 mg, 25%). MS (ESI): 552.5 (M+H)⁺.

Example 172

Preparation ofN-[N-methyl-N-(2-pyridinylmethoxycarbonyl-L-leucinyl]-N′-[2-[N-(2-methylpropyl-N-phenylamino]thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 116(a)-116(b), except substitutingN-methyl-N-(2-pyridinylmethoxycarbonyl)-L-leucine forN-(4-pyridinylmethoxycarbonyl)-L-leucine in step (b), the tide compoundwas prepared as a white solid solid (70 mg, 71%). MS (ESI): 553.4(M+H)⁺.

Example 173

Preparation ofN-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]-N′-(N-tert-butoxycarbonyl-L-leucinyl)hydrazide

Following the procedure of Example 112(a)-112(g), except substitutingN-tert-butoxycarbonyl-L-leucine forN-(4-pyridinylmethoxycarbonyl)-L-leucine in step (g), the title compoundwas prepared as a white solid (1.015 g, 94%). MS(ESI): 539.1 (M+H)⁺.

Example 174

Preparation ofN-(N-tert-butoxycarbonyl-L-leucinyl)-N′-[2-[N-(2-methylpropyl)-N-phenylamino]thiazolyl-4-ylcarbonyl]hydrazide

Following the procedure of Example 116(a)-116(b), except substitutingN-tert-butoxycarbonyl-L-leucine forN-(4-pyridinylmethoxycarbonyl)-L-leucine in step (b), the tide compoundwas prepared as a white solid (740 mg, 85%). MS (ESI): 504.4 (M+H)⁺.

Example 175

Preparation ofN-(N-tert-butoxycarbonyl-N-methyl-L-leucinyl)-N′-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 116(a)-116(b), except substitutingN-tert-butoxycarbonyl-N-methyl-L-leucine forN-(4-pyridinylmethoxycarbonyl)-L-leucine in step (b), the title compoundwas prepared as a white solid (610 mg, 69%). MS (ESI): 518.4 (M+H)⁺.

Example 176

Preparation ofN-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]-N′-(N-pyrazinecarbonyl-L-leucinyl)hydrazide

a)N-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]-N′-(L-leucinyl)hydrazide

Following the procedure of Example 144(b), except substitutingN-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]-N′-(N-tert-butoxycarbonyl-L-leucinyl)hydrazidefor(1S)-N-[2-[1-(N-tert-butoxycarbonyl-N-methylamino)-3-methylbutyl]thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide,the tide compound was prepared as a white powder (0.766 g, 93%).MS(ESI): 439.3 (M+H)⁺.

b)N-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]-N′-[N-(2-pyrazinylcarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 116(b), except substitutingN-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]-N′-(L-leucinyl)hydrazideforN-[2-[N-phenyl-N-(2-methyl-1-propyl)amino]thiazol-4-ylcarbonyl]hydrazideand pyrazinecarboxylic acid forN-(4-pyridinylmethoxycarbonyl)-L-leucine, the title compound wasprepared as a white solid (0.146 g, 94%). MS(ESI): 545.4 (M+H)⁺.

Example 177

Preparation ofN-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]-N′-(N-isonicotinoyl-L-leucinyl)hydrazide

Following the procedure of Example 176(a)-176(b), except substitutingisonicotinic acid for pyrazinecarboxylic acid in step (b), the titlecompound was prepared as a white solid (0.135 g, 87%). MS(ESI): 544.3(M+H)⁺.

Example 178

Preparation ofN-[2-(2-dibenzofuranyl)thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylymethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 112(a)-112(g), except substituting2-bromodibenzofuran for 2-benzyloxybromobenzene in step (d), the titlecompound was prepared as a white solid (0.079 g, 49%). MS(ESI): 558.3(M+H)⁺.

Example 179

Preparation ofN-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]-N′-(N-pyrazinecarbonyl-L-leucinyl)hydrazide

Following the procedure of Example 176(a)-176(b), except substitutingN-(N-tert-butoxycarbonyl-L-leucinyl)-N′-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]hydrazideforN-[2-(2-benzyloxyphenyl)thiazol-4ylcarbonyl]-N′-(N-tert-butoxycarbonyl-L-leucinyl)hydrazidein step (a), the title compound was prepared as a white solid (36 mg,27%). MS (ESI): 510.4 (M+H)⁺.

Example 180

Preparation ofN-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]-N′-(N-methyl-N-pyrazinecarbonyl-L-leucinyl)hydrazide

Following the procedure of Example 176(a)-176(b), except substitutingN-(N-tert-butoxycarbonyl-N-methyl-L-leucinyl)-N′-[2-[N-(2-methylpropyl)-N-phenylamino]thiazolylcarbonyl]hydrazideforN-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]-N′-(N-tert-butoxycarbonyl-L-leucinyl)hydrazidein step (a), the title compound was prepared as a white solid (70 mg,72%). MS (ESI): 524.4 (M+H)⁺.

Example 181

Preparation ofN-(N-isonicotinoyl-L-leucinyl)-N′-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 176(a)-176(b), except substitutingN-(N-tert-butoxycarbonyl-N-methyl-L-leucinyl)-N′-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]hydrazideforN-[2-(2-benzyloxyphenyl)thiazol-4ylcarbonyl]-N′-(N-tert-butoxycarbonyl-L-leucinyl)hydrazidein step (a) and isonicotinic acid for pyrazinecarboxylic aicd in step(b), the title compound was prepared as a white solid (28 mg, 22%). MS(ESI): 509.4 (M+H)⁺.

Example 182

Preparation ofN-(N-isonicotinoyl-N-methyl-L-leucinyl)-N′-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 176(a)-176(b), except substitutingN-(N-methyl-N-tert-butoxycarbonyl-N-methyl-L-leucinyl)-N′-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4ylcarbonyl]hydrazidefor N-[2-(2-benzyloxyphenyl)thiazol-4ylcarbonyl]-N′-(N-tert-butoxycarbonyl-L-leucinyl)hydrazide in step (a)and isonicotinic acid for pyrazinecarboxylic aicd in step (b), the titlecompound was prepared as a white solid (117 mg, 93%). MS (ESI): 523.4(M+H)⁺.

Example 183

Preparation ofN-[N-(4-imidazolylacetyl)-L-leucinyl]-N′-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 176(a)-176(b), except substitutingN-(N-tert-butoxycarbonyl-N-methyl-L-leucinyl)-N′-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]hydrazideforN-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]-N′-(N-tert-butoxycarbonyl-L-leucinyl)hydrazidein step (a) and 4-imidazolylacetic acid for pyrazinecarboxylic aicd instep (b), the title compound was prepared as a white solid (60 mg, 53%).¹HNMR (400 MHz, CDCl₃) δ 7.62-7.23 (m, 8H), 6.81 (s, 1H), 4.724.66 (m,1H), 3.75 (d, 1H), 3.55 (d, 2H), 1.96-1.93 (m, 2H), 1.76-1.54 (m, 3H),0.96-0.84 (m, 12H).

Example 184

Preparation ofN-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]-N′-(N-picolinoyl-L-leucinyl)hydrazide

Following the procedure of Example 176(a)-176(b), except substitutingN-(N-tert-butoxycarbonyl-N-methyl-L-leucinyl)-N′-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]hydrazideforN-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]-N′-(N-tert-butoxycarbonyl-L-leucinyl)hydrazidein step (a) and picolinic acid for pyrazinecarboxylic aicd in step (b),the title compound was prepared as a white solid (50 mg, 44%). MS (ESI):509.5 (M+H)⁺.

Example 185

Preparation of N-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]methyl[N-(4-pyridinylmethoxycarbonyl)-L-leucinamide

a) 2-(2-benzyloxyphenylthiazole-4-carboxylic Acid

Following the procedure of Example 105(b), except substituting ethyl2-(2-benzyloxyphenyl)thiazole-4-carboxylate for methyl3-(4-pyridinyllmethoxy)benzoate, the title compound was prepared as awhite solid (0.361 g, 90%). MS(ESI): 312.2 (M+H)⁺.

b) 2-(2-benzyloxyphenyl)thiazol-4-yl bromomethyl Ketone

Following the procedure of Example 103(a), except substituting2-(2-benzyloxyphenylthiazole-4-carboxylic acid forN-benzyloxycarbonyl-L-Leucinyl-L-Leucine, the title compound wasprepared as a white solid (0.327 g, 73%). MS(ESI): 388.2 (M+H)⁺.

c) 2-(2-benzyloxyphenyl)thiazol-4-yl Azidomethyl Ketone

A solution of the compound of Example 185(b), (0.319 g, 0.822 mmol),sodium azide (0.064 g. 0.987 mmol), and potassium fluoride (0.072 g,1.23 mmol) in DMF (6 mL) was stirred at room temperature for 16 h. Thesolution was then diluted with ethyl acetate and washed successivelywith water, saturated aqueous sodium hydrogen carbonate, and brine. Theorganic layer was dried (MgSO₄), filtered and concentrated. The residuewas purified by column chromatography (silica gel, ethyl acetate/hexane)to yield the title compound as a white solid (0.087 g, 30%). MS(ESI):373.3 (M+Na)⁺.

d) 2-azido-1-[2-(2-benzyloxyphenyl)thiazol-4-yl]-1-hydroxyethane

To a stirring solution of the compound of Example 185(c) (0.087 g, 0.249mmol) in THF (1 mL) at 0° C., was added sodium borohydride (0.031 g,0.820 mmol) slowly. After 20 min the mixture was diluted with ethylacetate and washed with water then brine. The organic layer was dried(MgSO₄), filtered and concentrated to yield the title compound as awhite solid (0.084 g, 96%). 1 (400 MHz, CDCl₃) δ 8.41 (m, 1H), 7.50 (m,2H), 7.38 (m, 4H), 7.11 (m, 3H), 5.31 (s, 2H), 5.08 (t, 1H), 3.69 (m,2H), 3.58 (s b, 1H).

e) 2-amino-1-[2-(2-benzyloxyphenyl)thiazol-4-yl]-1-hydroxyethane

To a stirring solution of the compound of Example 185(d) (0.084 g, 0.239mmol) in methanol (2 mL) was added stannous chloride dihydrate (0.108 g,0.478 mmol). After stirring at room temperature for 16 h, the mixturewas diluted with ethyl acetate and washed successively with water,saturated aqueous NaHCO₃, and brine. The organic layer was dried(MgSO₄), filtered and concentrated. The residue was purified by columnchromatography (silica gel, methanol/dichloromethane) to yield the titlecompound as a white solid (0.07 5 g, 96%). MS(ESI): 327.3 (M+H)⁺.

f)1-[2-(2-benzyloxyphenyl)thiazol-4-yl]-1-hydroxy-2-(4-pyridinylmethoxycarbonyl-L-leucinylamino)ethane

Following the procedure of Example 116(b), except substituting2-amino-1-[2-(2-benzyloxyphenyl)thiazol-4-yl]-1-hydroxyethane forN-[2-[N-phenyl-N-(2methyl-1-propyl)amino]thiazol-4-ylcarbonyl]hydrazide, the title compoundwas prepared as a white solid (0.075 g, 57%). MS(ESI): 575.4 (M+H)⁺.

g) N-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]methyl[N-(4-pyridinylmethoxycarbonyl)-L-leucinamide

To a stirring solution of the compound of Example 185(f) (0.075 g, 0.131mmol) in dichloromethane (1 mL) was added MnO₂ (0.300 g, 3.45 mmol).After stirring at room temperature for 24 h, the mixture was filteredand the filtrate was concentrated. The residue was purified by columnchromatography (silica gel, methanol/dichloromethane) to yield the tidecompound as a pale yellow solid (0.017 g, 23%). MS(ESI): 573.4 (M+H)⁺.

Example 186

Preparation ofN-[2-[N-methyl-N-(2-methylpropyl)amino]thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

a) N-benzoyl-N′-methyl-N′-(2-methylpropyl)thiourea

To a stirring solution of N-methylisobutylamine (3.21 g, 36.8 mmol, 4.45mL) in 40 mL of CHCl₃ was added benzoyl isothiocyanate (6.0 g, 36.8mmol, 4.95 mL) After stirring for 45 min, the solution was concentratedto yield the title compound as a pale yellow solid (9.22 g, 100%). ¹HNMR(400 MHz, CDCl₃; 2:1 mixture of rotamers) δ 7.86 (d, 2H), 7.60 (t, 1H),7.50 (t, 2H), 3.90 (d, 2H), 3.44 (s, 3H), 3.41 (d, 2H), 3.27 (s, 3H),2.35-2.32 (m, 1H), 2.13 (m, 1H), 1.06 (d, 6H), 0.90 (d, 6H).

b) N-methyl-N-(2-methylpropyl)thiourea

The compound of Example 186(a) (9.22 g. 36.8 mmol) was suspended in 80mL of 1:1 methanol/water and solid potassium carbonate (15.27 g, 110mmol) was added. The mixture was heated at reflux for 48 h, then cooledand partitioned between ethyl acetate and water. The aqueous layer wasextracted with ethyl acetate (2×). The combined organic layers werewashed with saturated brine, dried (MgSO₄), filtered and concentrated toprovide the title compound as a pale yellow crystalline solid (4.82 g,89%). MS(ESI): 147.0 (M+H)⁺.

c)N-[2-[N-methyl-N-(2-methylpropyl)amino]thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 113(d)-113(f), except substitutingN-methyl-N-(2-methylpropyl)thiourea forN-(4-phenylphenyl)-N-(2-methyl-1-propyl)thiourea in step (d), the titlecompound was prepared as a white solid (202 mg, 97%). MS(ESI): 477.4(M+H)⁺.

Example 187

Preparation ofN-(N-methyl-N-picolinoyl-L-leucinyl)-N′-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 176(a)-176(b), except substitutingN-(N-methyl-N-tert-butoxycarbonyl-N-methyl-L-leucinyl)-N′-[2-[N-(2-methylpropyl)-N-phenylamino]thiazolylcarbonyl]hydrazideforN-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]-N′-(N-tert-butoxycarbonyl-L-leucinyl)hydrazidein step (a) and picolinic acid for pyrazinecarboxylic aicd in step (b),the title compound was prepared as a white solid (30 mg, 41%). MS (ESI):523.5 (M+H)⁺.

Example 188

Preparation ofN-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]-N′-[N-(2-pyridinesulfonyl)-L-leucinyl]hydrazide

a) 2-pyridinesulfonylchloride

Through a stirring solution of 2-mercaptopyridine (2.235 g, 20 mmol) inwater (7.5 mL) and concentrated HCl (26 mL) at 0° C. was bubbled Cl₂.After 30 min, 75 mL of ice water was added and extracted with cold ether(2×75 mL). The organic layers were combined and washed successively withcold 10% aqueous NaHCO₃, and cold brine. The organic layer was dried(MgSO₄), filtered and concentrated to yield the title compound as aclear oil. (3.1 g, 87%).

b)N-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]-N′-[N-(2-pyridinesulfonyl)-L-leucinyl]hydrazide

To a stirring solution of the compound of Example 176(a) (0.125 g, 0.285mmol), and the compound of Example 188(a) (0.101 g, 0.571 mmol) indichloromethane (2 mL) was added N-methylmorpholine (0.057 g, 0.571mmol). After stirring at room temperature for 10 min the solution wasdiluted with ethyl acetate and washed successively with water and brine.The organic layer was dried (MgSO₄), filtered and concentrated. Theresidue was purified by column chromatography (silica gel, ethylacetate/hexane) to yield the title compound as a pale yellow solid(0.100 g, 61%). MS(ESI): 580.2 (M+H)⁺.

Example 189

Preparation ofN-[2-[N-(2-methylpropyl)N-phenylamino]thiazol-4-ylcarbonyl]-N′-[N-(2-pyridinesulfonyl)-L-leucinyl]hydrazide

Following the procedure of Example 188(b), except substitutingN-(L-leucinyl)-N′-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]hydrazideforN-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]-N′-(L-leucinyl)hydrazide,the title compound was prepared as an orange solid (56 mg, 48%). MS(ESI): 545.3 (M+H)⁺.

Example 190

Preparation ofN-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]-N′-[N-methyl-N-(2-pyridinesulfonyl)-L-leucinyl]hydrazide

Following the procedure of Example 188(b), except substitutingN-(N-methyl-L-leucinyl)-N′-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]hydrazideforN-[2-(2-benzyloxyphenyl)thiazol-4ylcarbonyl]-N′-(L-leucinyl)hydrazide,the title compound was prepared as an orange solid (53 mg, 40%). MS(ESI): 559.3 (M+H)⁺.

Example 191

Preparation ofN-[2-[N-methyl-N-(2-methylpropyl)amino]thiazol-4-ylcarbonyl]-N′-[N-(3-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 186(a)-186(c), except substitutingN-(3-pyridinylmethoxycarbonyl)-L-leucine forN-(4pyridinylmethoxycarbonyl)-L-leucine in step (c), the title compoundwas prepared as a white solid (138 mg, 66%). MS (ESI): 477.4 (M+H)⁺.

Example 192

Preparation ofN-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]-N′-[N-methyl-N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 116(a)-116(b), except substitutingN-methyl-N-(4-pyridinylmethoxycarbonyl)-L-leucine forN-(4-pyridinylmethoxycarbonyl)-L-leucine in step (b), the tide compoundwas prepared as a white solid solid (74 mg, 41%). MS (ESI): 553.4(M+H)⁺.

Example 193

Preparation ofN-[2-[N-(2-methylpropyl)-N-phenylamino]thiazol-4-ylcarbonyl]-N-[N-methyl-N-(3-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 116(a)-116(b), except substitutingN-methyl-N-(3-pyridinylmethoxycarbonyl)-L-leucine forN-(4-pyridinylmethoxycarbonyl)-L-leucine in step (b), the title compoundwas prepared as a white solid solid (50 mg, 38%). MS (ESI): 553.4(M+H)⁺.

Example 194

Preparation ofN-[2-(2-benzyloxyphenyl)thiazol-4-ylcarbonyl]-N′-[N-methyl-N-(3-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 112(a)-112(g), except substitutingN-methyl-N-(4-pyridinylmethoxycarbonyl)-L-leucine forN-(4-pyridinylmethoxycarbonyl)-L-leucine in step (g), the title compoundwas prepared as a white solid (0.028 g, 15%). MS(ESI): 588.4 (M+H)⁺.

Example 195

Preparation ofN-[N-methyl-N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]-N′-[2-(1-naphthyl)thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 112(a)-112(g), except substituting1-naphthyl boronic acid for 2-benzyloxyphenyl boronic acid in step (e)and N-methyl-N-(4-pyridinylmethoxycarbonyl)-L-leucine forN-(4-pyridinylmethoxycarbonyl)-L-leucine in step (g), the title compoundwas prepared as a white solid (0.072 g, 36%). MS(ESI): 532.4 (M+H)⁺.

Example 196

Preparation ofN-[2-[N,N-(bis)-2-methylpropyl)amino]thiazol-4-ylcarbonyl]-N-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 186(a)-186(c), except substitutingN,N-diisobutylamine for N-methylisobutylamine in step (a), the titlecompound was prepared as a yellow solid (60 mg, 39%). MS (ESI): 519.5(M+H)⁺.

Example 197

Preparation ofN-(N-benzyloxycarbonyl-L-leucinyl)-N′-[2-[N,N-(bis)-2-methylpropyl)amino]thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 186(a)-186(c), except substitutingN,N-diisobutylamine for N-methylisobutylamine in step (a) andN-benzyloxycarbonyl-L-leucine forN-(4-pyridinylmethoxycarbonyl)-L-leucine in the final step, the titlecompound was prepared as a yellow solid (131 mg, 69%). MS (ESI): 518.4(M+H)⁺.

Example 198

Preparation ofN-[2-(4-morpholino)thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 186(a)-186(c), except substitutingmorpholine for N-methylisobutylamine in step (a), the title compound wasprepared as a white solid (45 mg, 31%). MS (ESI): 477.3 (M+H)⁺.

Example 199

Preparation ofN-[2-[2-(4-pyridinylmethoxy)phenyl]thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

a) 2-methoxymethoxybromobenzene

To a stirring suspension of sodium hydride (1.2 g, 52.1 mmol, 60%dispersion in mineral oil) in DMF (75 mL) at 0° C. was added2-bromophenol (6.0 g, 34.7 mmol) dropwise. After stirring for 20 min,bromomethyl methyl ether (4.3 g, 34.7 mmol) was added. After stirringfor 16 h at room temperature, the mixture was poured into water (250 mL)and extracted with hexane. The organic layer was washed with brine,dried (MgSO₄), filtered and concentrated. The residue was purified bycolumn chromatography (silica gel, hexane) to yield the title compoundas a colorless oil (4.0 g, 53%). ¹HNMR (400 MHz, CDCl₃) δ 7.55 (d, 1H),7.28 (t, 1H), 7.16 (d, 1H), 6.91 (t, 1H), 5.25 (s, 2H), 3.54 (s, 3H).

b) ethyl 2-(2-methoxymethoxyphenyl)thiazole-4-carboxylate

Following the procedure of Example 112(a)-112(b) and 112(d)-112(e),except substituting 2-methoxymethoxybromobenzene for2-benzyloxybromobenzene in step (d), the title compound was prepared.MS(ESI): 294.3 (M+H)⁺.

c) ethyl 2-(2-hydroxyphenyl)thiazole-4-carboxylate

To a stirring solution of the compound of Example 199(b) (0.839 g, 2.86mmol) in ethanol (25 mL) was added 10 drops of concentrated hydrochloricacid. After stirring at reflux for 2 h the solution was concentratedthen redissolved in ethyl acetate. The solution was washed successivelywith saturated sodium bicarbonate, and brine, dried (MgSO₄), filteredand concentrated to yield the title compound as a pale yellow solid(0.674 g, 95%). MS(ESI): 250.2 (M+H)⁺.

d) ethyl 2-[2-(4-pyridylmethoxy)phenyl]thiazole-4-carboxylate

To a stirring solution of the compound of Example 199(c) (0.125 g, 0.502mmol), 4-pyridylcarbinol (0.071 g, 0.653 mmol), and triphenylphosphine(0.171 g, 0.653 mmol) in THF (5 mL) at 0° C. was added diisopropylazodicarboxylate (0.132 g, 0.653 mmol) dropwise. After stirring at roomtemperature for 16 h, the solution was concentrated and purified bycolumn chromatography (silica gel, ethyl acetate/hexane) to yield thetitle compound as a white solid (0.100 g, 59%). MS(ESI): 341.3 (M+H)⁺.

e)N-[2-[2-(4-pyridinylmethoxy)phenyl]thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 112(f)-112(g), except substitutingethyl 2-[2-(4-pyridylmethoxy)phenyl]thiazole-4-carboxylate for ethyl2-(2-benzyloxyphenyl)thiazole-4-carboxylate in step (f), the titlecompound was prepared as a white solid (146 mg, 83%). MS(ESI): 575.4(M+H)⁺.

Example 200

Preparation ofN-[2-(2-naphthyl)thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 112(a)-112(g), except substituting2-naphthylboronic acid for 2-benzyloxyphenyl boronic acid in step (e),the title compound was prepared as a white solid (226 mg, 75%). MS(ESI): 518.4 (M+H)⁺.

Example 201

Preparation ofN-[2-[N,N-(bis)-2-methylpropyl)amino]thiazol-4ylcarbonyl]-N′-[N-methyl-N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 186(a)-186(c), except substitutingN,N-diisobutylamine for N-methylisobutylamine in step (a) andN-methyl-N-(4-pyridinylmethoxycarbonyl)-L-leucine forN-(4-pyridinylmethoxycarbonyl)-L-leucine in the final step, the titlecompound was prepared as a yellow solid (30 mg, 25%). MS (ESI)-533.3(M+H)⁺.

Example 202

Preparation ofN-(N-benzyloxycarbonyl-L-leucinyl)-N′-[2-(4-morpholino)thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 186(a)-186(c), except substitutingmorpholine for N-methylisobutylamine in step (a) andN-benzyloxycarbonyl-L-leucine forN-(4-pyridinylmethoxycarbonyl)-L-leucine in the final step, the titlecompound was prepared as a white solid (115 mg, 67%). MS (ESI): 576.4(M+H)⁺.

Example 203

Preparation ofN-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]-N′-[2-(4-thiomorpholino)thiazol-4ylcarbonyl]hydrazide

Following the procedure of Example 186(a)-186(c), except substitutingthiomorpholine for N-methylisobutylamine in step (a), the title compoundwas prepared as a white solid (35 mg, 20%). MS (ESI): 493.4 (M+H)⁺.

Example 204

Preparation ofN-(N-benzyloxycarbonyl-L-leucinyl)-N′-[2-(4-thiomorpholino)thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 186(a)-186(c), except substitutingthiomorpholine for N-methylisobutylamine in step (a) andN-benzyloxycarbonyl-L-leucine forN-(4-pyridinylmethoxycarbonyl)-L-leucine in the final step, the titlecompound was prepared as a white solid (20 mg, 20%). MS (ESI): 492.3(M+H)⁺.

Example 205

Preparation ofN-[2-(2,3-ethylenedioxy-4-methoxyphenyl)thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 112(a)-112(g), except substituting2,3-ethylenedioxy-4-methoxybromobenzene for 2-benzyloxybromobenzene instep (d), the title compound was prepared as a white solid (31 mg, 26%).MS (ESI): 556.4 (M+H)⁺.

Example 206

Preparation ofN-[2-[N,N-(bis)-2-methylpropyl)amino]thiazol-4-ylcarbonyl]-N′-[N-methyl-N-(3-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 186(a)-186(c), except substitutingN,N-diisobutylamine for N-methylisobutylamine in step (a) andN-methyl-N-(3-pyridinylmethoxycarbonyl)-L-leucine forN-(4-pyridinylmethoxycarbonyl)-L-leucine in the final step, the titlecompound was prepared as a yellow solid (30 mg, 30%). MS (ESI): 533.5(M+H)⁺.

Example 207

Preparation ofN-[2-(N-cyclopropylmethyl-N-propylamino)thiazol-4-ylcarbonyl]-N′-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]hydrazide

Following the procedure of Example 186(a)-186(c), except substitutingN-cyclopropylmethylpropylamine for N-methylisobutylamine in step (a),the title compound was prepared as a yellow solid (60 mg, 23%). MS(ESI): 503.3 (M+H)⁺.

Example 208

Preparation ofN-[N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]-N′-[2-(8-quinolyl)thiazol-4-ylcarbonyl]hydrazide

Following the procedure of Example 112(a)-112(g), except substituting8-bromoquinoline for 2-benzyloxybromobenzene in step (d), the titlecompound was prepared as a white solid (134 mg, 56%). MS (ESI): 519.3(M+H)⁺.

Example 209

Preparation ofN-[N-methyl-N-(4-pyridinylmethoxycarbonyl)-L-leucinyl]-N′-[2-(8-quinolyl)thiazol-4-ylcarbonyl]hydrazideFollowing the procedure of Example 112(a)-112(g), except substituting8-bromoquinoline for 2-benzyloxybromobenzene in step (d) andN-methyl-N-(4-pyridinylmethoxycarbonyl)-L-leucine forN-(4-pyridinylmethoxycarbonyl)-L-leucine in step (g), the title compoundwas prepared as a white solid (53 mg, 22%). MS (ESI): 533.3 (M+H)⁺.

Example 210

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-(4-biphenyl-sulfonyl)-amino-propan-2-one

a) 4-biphenyl Sulfonyl Chloride

4-Biphenyl sulfonic acid (2.4 g, 10 mmol) was heated to 100 C withphosphorus pentachloride (2.1 g, 10 mmol) overnight. The reaction wascooled to RT, diluted with water, filtered and washed with water. Thesolid was then triturated with EtOAc-ether and the beige solid was usedin the next reaction without further purification.

b)1-N-(N-Cbz-leucinyl)-amino-3-N-(4-biphenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 51(a), except substituting“4-biphenyl sulfonyl chloride” for “4-(3-Chloro-2-cyano-phenoxy)-phenylsulfonyl chloride”, the title compound was prepared: MS(ES) M−H⁺=550.

Example 211

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-(3-biphenyl-sulfonyl)-amino-propan-2-one

a) 3-biphenyl-sulfonyl Chloride

3-Biphenyl bromide (9.3 g, 40 mmol) was dissolved in THF (40 ml) and aGrignard reagent was prepared in standard fashion with magnesium powder(1.2 g, 50 mmol). The reaction was cannulated into a solution ofsulfuryl chloride (10.5 g, 6.4 ml, 80 mmol) in hexanes (25 ml) and wasstrirred at RT for 2 h. The reaction was quenched with ice-water,extracted with ether, dried with magnesium sulfate, filtered,concentrated, and was used in the next reaction without furtherpurification.

b)1-N-(N-Cbz-leucinyl)-amino-3-N-(3-biphenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 51(a), except substituting“3-biphenyl sulfonyl chloride” for “4-(3-Chloro-2-cyano-phenoxy)-phenylsulfonyl chloride”, the title compound was prepared: MS(ES) M−H⁺=550.

Example 212

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-(2-benzyloxy-phenyl-sulfonyl)-amino-propan-2-one

a) 2-benzyloxy-phenyl-sulfonyl Chloride

Following the procedure of Example 211(a), except substituting“2-benzyloxy-phenyl bromide” for “3-biphenyl bromide”, the titlecompound was prepared and was used in the next step without furtherpurification.

b)1-N-(N-Cbz-leucinyl)-amino-3-N-(2-benzyloxy-phenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 51(a), except substituting“2-benzyloxy phenyl sulfonyl chloride” for“4-(3-Chloro-2-cyano-phenoxy)-phenyl sulfonyl chloride”, the tidecompound was prepared: MS(ES) M+H⁺=581, M+Na⁺=604, 2M+Na⁺=1185.

Example 213

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-(4-phenoxy-phenyl-sulfonyl)-amino-propan-2-one

a) 4-phenoxy-phenyl-sulfonyl Chloride

Following the procedure of Example 211(a), except substituting“4-phenoxy phenyl bromide” for “3-biphenyl bromide”, the title compoundwas prepared and was used in the next step without further purification.

b)1-N-(N-Cbz-leucinyl)-amino-3-N-(4-phenoxy-phenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 51(a), except substituting“4-phenoxy-phenyl-sulfonyl chloride” for“4-(3-Chloro-2-cyano-phenoxy)-phenyl sulfonyl chloride”, the titlecompound was prepared: MS(ES) M+H⁺=568, M+Na⁺=590.

Example 214

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-(2-dibenzofuran-sulfonyl)-amino-propan-2-one

a) 4-phenoxy-phenyl-sulfonyl Chloride

Following the procedure of Example 210(a), except substituting“2-dibenzofuran-sulfonic acid” for “4-biphenyl-sulfonic acid”, the titlecompound was prepared and was used in the next step without furtherpurification.

b)1-N-(N-Cbz-leucinyl)-amino-3-N-(2-dibenzofuran-sulfonyl)-amino-propan-2-one

Following the procedure of Example 51(a), except substituting “4-phenoxyphenyl sulfonyl chloride” for “4-(3-Chloro-2-cyano-phenoxy)-phenylsulfonyl chloride”, the title compound was prepared: MS(ES) M+H⁺=566,M+Na⁺=588.

Example 215

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-(3,4dimethoxy-phenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 51, except substituting“3,4-dimethoxy-phenyl-sulfonyl chloride” for“4-(3-Chloro-2-cyano-phenoxy)-phenyl sulfonyl chloride”, the titlecompound was prepared: MS(ES) M+H⁺=536, M+NH₄ ⁺=553.

Example 216

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-(2,5-dichlorothiophene-3-sulfonyl)-amino-propan-2-one

Following the procedure of Example 51, except substituting“2,5-dichlorothiophene-3-sulfonyl chloride” for“4-(3-Chloro-2-cyano-phenoxy)-phenyl sulfonyl chloride”, the tidecompound was prepared: MS(ES) M+NH₄ ⁺=567, 2M+H⁺=1101.

Example 217

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-(phenyl-sulfone-5-thiophene-2-sulfonyl)-amino-propan-2-one

Following the procedure of Example 51, except substituting “phenylsulfone-5-thiophene-2-sulfonyl chloride” for“4-(3-Chloro-2-cyano-phenoxy)-phenyl sulfonyl chloride”, the titlecompound was prepared: MS(ES) M+H⁺=622.

Example 218

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-(8-quinoline-sulfonyl)-amino-propan-2-one

Following the procedure of Example 51, except substituting“8-quinoline-sulfonyl chloride” for “4-(3-Chloro-2-cyano-phenoxy)-phenylsulfonyl chloride”, the title compound was prepared: MS(ES) M+H³⁰ =527.

Example 219

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-(2-pyridyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 51, except substituting“2-pyridyl-sulfonyl chloride” (as described in J. Org. Chem. 1989, 54,392) for “4-(3-Chloro-2-cyano-phenoxy)-phenyl sulfonyl chloride”, thetitle compound was prepared: MS(ES) M+H⁺=477, M+Na⁺=499.

Example 220

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-(2-pyridyl-sulfonyl)-amino-propan-2-one

a)1-N-(N-Cbz-leucinyl)-amino-3-N-(4-phenoxy-phenyl-sulfonyl)-amino-propan-2-ol

1,3-Diamino propan-2-ol (6.75 g, 75 mmol) was dissolved in DMF (100 ml)and Cbz-leucine (20 g, 75.5 mmol), HOBT-hydrate (11 g, 81.5 mmol), andEDCI (15.5 g, 81.2 mmol) were added. The reaction was stirred overnightat RT. A portion of the reaction mixture (30 ml) was concentrated invacuo, then ether (50 ml) and MeOH (30 ml) were added. A 1N solution ofhydrochloric acid in ether was added (1 M, 30 ml) and a white gumformed, which was washed several times with ether. MeOH-acetone wereadded and heated until the gum became a white solid. The white solid wasdissolved in DMF (25 ml) and DIEA (5 ml), then 4-phenoxy phenyl sulfonylchloride was added. The reaction was stirred for 2 h, concentrated invacuo, then chromatographed (silica gel, 1:1 EtOAc: hexanes) to providethe desired product as a white solid.

b) Leucinyl-amino-3-N-(4-phenoxy phenyl sulfonyl)-amino-propan-2-ol

1-N-(Cbz-leucinyl)-amino-3-N-(4-phenoxy-phenyl-sulfonyl)-amino-propan-2-ol(1.0 g, 1.8 mmol) was dissolved in EtOH (30 ml), then 10% Pd/C (0.22 g)was added followed by 6N hydrochloric acid (2.5 ml), and the reactionwas stirred under a baloon of hydrogen gas for 4 h at RT. The reactionmixture was filtered, concentrated, and azeotroped with toluene toprovide a white glass which was used in the next reaction withoutfurther purification.

c) 1-N-(N-4-pyridylacetyl-leucinyl)-amino-3-N-(4phenoxy-phenyl-sulfonyl)-amino-propan-2-ol

Leucinyl-amino-3-N-(4-phenoxy phenyl sulfonyl)-amino-propan-2-ol (0.36g, 0.76 mmol) was dissolved in DMF (5 ml), then NMM (0.45 ml, 4 mmol)was added followed by 4-pyridyl acetic acid (0. 13 g, 0.75 mmol) andHBTU (0.29 g, 0.76 mmol) and the reaction was stirred at RT overnight.The reaction mixture was concentrated in vacuo, then chromatographed(silica gel, 5% MeOH: methylene chloride) to provide the desired productas a white solid (90 mg, MS(ES): M+H⁺=555.

d) 1-N-(N-4-pyridylAcetyl-leucinyl)-amino-3-N-(4phenoxy-phenyl-sulfonyl)-amino-propan-2-one

1-N-(N-4-pyridyl-acetyl-leucinyl)-amino-3-N-(4-phenoxy-phenyl-sulfonyl)-amino-propan-2-ol(45 mg, 0.08 mmol) was dissolved in acetone (5 ml), then 1N hydrochloricacid (2 ml) was added. The reaction was concentrated in vacuo, thenredissolved in acetone. Jones reagent (1.5 M, several drops) was addedand the reaction mixture was stirred for 6 h at RT. Isopropanol (0.5 ml)was added and the reaction mixture was concentrated in vacuo. Thereaction was diluted with pH 7 buffer and then was extracted with EtOAc,dried with magnesium sulfate, filtered, concentrated in vacuo, thenchromatographed (silica gel, 5% MeOH-methylene chloride) to give thedesired product as a white solid (27 mg, 50%): MS(ES): M+H⁺=553.

Example 221

Preparation of1-N-(N-2-pyridyl-sulfonyl-leucinyl)-amino-3-N-(4-phenoxy-phenyl-sulfonyl)amino-propan-2-one

Following the procedure of Example 220(a)-(d), except substituting“2-pyridyl-sulfonyl chloride” (as described in J. Org. Chem. 1989, 54,392) for “4-pyridyl-acetic acid and HBTU”, the title compound wasprepared: MS(ES) M+H⁺=475, M+Na⁺=497, 2M+Na⁺=1171.

Example 222

Preparation of1-N-(N-morpholino-carbonyl-leucinyl)-amino-3-N-(4-phenoxy-phenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 220(a)-(d), except substituting“N-morpholino-carbonyl chloride” for “4-pyridyl acetic acid and HBTU”,the title compound was prepared: MS(ES) M+H⁺=547, M+Na⁺=569,2M+Na⁺=1115.

Example 223

Preparation of1-N-(N-4-pyridyl-carbonyl-leucinyl)-amino-3-N-(4-phenoxy-phenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 220(a)-(d), except substituting“4-pyridyl-carboxylic acid” for “4pyridyl acetic acid”, the tidecompound was prepared: MS(ES) M+H⁺=539.

Example 224

Preparation of1-N-(N-acetyl-leucinyl)-amino-3-N-(4-phenoxy-phenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 220(a)-(d), except substituting“acetyl chloride” for “4-pyridyl-acetic acid and HBTU”, the tidecompound was prepared: MS(ES) M+H⁺=476, M+Na⁺=498, 2M+Na⁺=973.

Example 225

Preparation of 1-N-(N-imidazoleacetyl-leucinyl)-amino-3-N-(4-phenoxy-phenyl-sulfonyl)-amino-propan-2one

Following the procedure of Example 220(a)-(d), except substituting“imidazole acetic acid” for “4-pyridyl acetic acid”, the title compoundwas prepared: MS(ES) M+H⁺=542.

Example 226

Preparation of 1-N-(N-4-carboxymethylbenzoyl-leucinyl)-amino-3-N-(4phenoxy-phenyl-sulfonyl)-amino-propan-2one

Following the procedure of Example 220(a)-(d), except substituting“4-carboxymethyl benzoic acid” for “4-pyridyl acetic acid”, the titlecompound was prepared: MS(ES) M+H⁺=596, M+Na⁺=618, 2M+Na⁺=1213.

Example 227

Preparation of 1-N-(N-(N,N-dimethylglycinyl)-leucinyl)-amino-3-N-(4-phenoxy-phenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 220(a)-(d), except substituting“N,N-dimethyl glycine” for “4-pyridyl acetic acid”, the title compoundwas prepared: MS(ES) M+H⁺=519.

Example 228

Preparation of1-quinoline-sulfonamide-leucinyl)-(4-phenoxy-phenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 220(a)-(d), except substituting“8-quinoline sulfonyl chloride” for “4-pyridyl-acetic acid and HBTU”,the title compound was prepared: MS(ES) M+H⁺=625.

Example 229

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-(8-quinoline-carbonyl)-amino-propan-2-one

1,3-Diamino propan-2-ol (6.75 g, 75 mmol) was dissolved in DMF (100 ml)and Cbz-leucine (20 g, 75.5 mmol), HOBT-hydrate (11 g, 81.5 mmol), andEDCI (15.5 g, 81.2 mmol) was added. The reaction was stirred overnightat RT. A portion of the reaction mixture (30 ml) was concentrated invacuo, then ether (50 ml) and MeOH (30 ml) were added. A 1N solution ofhydrochloric acid in ether was added (1 M, 30 ml) and a white gumformed, which was washed several times with ether. MeOH-acetone wereadded and heated until the gum became a white solid. The white solid(0.44 g, 1.11 mmol) was dissolved in DMF (3 ml) and NMM (0.33 ml, 0.3mmol), then 8-quinoline carboxylic acid (0.17 g, 1.0 mmol), and HBTU(0.38 g, 1.0 mmol) were added and the reaction was stirred at RTovernight. The reaction mixture was concentrated in vacuo, thenchromatographed (silica gel, 7:2 EtOAc: hexanes). The solid was thendissolved in acetone (5 ml), then IN hydrochloric acid (2 ml) was added.The reaction was concentrated in vacuo, then redissolved in acetone.Jones reagent (1.5 M, several drops) was added and the reaction mixturewas stirred for 6 h at RT. Isopropanol (0.5 ml) was added and thereaction mixture was concentrated in vacuo. The reaction was dilutedwith pH 7 buffer and then was extracted with EtOAc, dried with magnesiumsulfate, filtered, concentrated in vacuo, then chromatographed (silicagel, 5% MeOH-methylene chloride) to give the desired product as a whitesolid (23 mg, 41%): MS(ES): M+H⁺=491.

Example 230

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-(6quinoline-carbonyl)-amino-propan-2-one

Following the procedure of Example 229, except substituting“6quinoline-carboxylic acid” for “8-quinoline carboxylic acid”, thetitle compound was prepared: MS(ES) M+H⁺=491.

Example 231

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-(2-(4-biphenyl)-4-methyl-propanamide)-propan-2-one

Following the procedure of Example 229, except substituting“2-isobutyl-4-biphenyl acetic acid” for “8-quinoline carboxylic acid”,the title compound was prepared: MS(ES) M+H⁺=586, M+Na⁺=608,2M+Na⁺=1193.

Example 232

Preparation of 1-N-(N-Cbz-leucinyl)-amino-3-N-(N-pyridyl-methyleneoxycarbonyl-leucinyl)-amino-propan-2-one

Following the procedure of Example 229, except substituting “4-pyridylmethyleneoxy carbonyl leucine” for “8-quinoline-carboxylic acid”,thetitle compound was prepared: MS(ES) M+H⁺=584.

Example 233

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-(benzoyl)-amino-propan-2-one

Following the procedure of Example 229, except substituting “benzoylchloride” for “8-quinoline carboxylic acid and HBTU”, the title compoundwas prepared: MS(ES) M+H⁺=440, M+Na⁺=462, 2M+Na⁺=901.

Example 234

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-(2,4-dimethyl-3-sulfonyl)-amino-propan-2-one

Following the procedure of Example 51, except substituting“2,4-dimethyl-3-sulfonyl chloride” for“4(3-Chloro-2-cyano-phenoxy)-phenyl sulfonyl chloride”, the titlecompound was prepared: MS(ES) M+H⁺=494.

Example 235

Preparation of 1-N-(N-Cbz-leucinyl)-amino-3-N-(1,3-dimethyl-5-chloro-pyrazole-4-sulfonyl)-amino-propan-2-one

Following the procedure of Example 51, except substituting“1,3-dimethyl-5-chloro-pyrazole-4-sulfonyl chloride” for“4-(3-Chloro-2-cyano-phenoxy)-phenyl sulfonyl chloride”, the titlecompound was prepared: MS(ES) M+H⁺=494.

Example 236

Preparation of 1-N-(N-4-pyridyl-methyleneoxyCarbonyl-leucinyl)-amino-3-N-(4-phenoxy-phenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 213(a), except substituting“4-pyridyl-methyleneoxy carbonyl-leucine” for “Cbz-leucine”, the titlecompound was prepared: MS(ES) M+H⁺=569.

Example 237

Preparation of 1-N-(N-3-pyridyl-methyleneoxyCarbonyl-leucinyl)-amino-3-N-(4-phenoxy-phenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 213(a), except substituting“3-pyridyl-methyleneoxy carbonyl-leucine” for “Cbz-leucine”, the titlecompound was prepared: MS(ES) M+H⁺=569.

Example 238

Preparation of1-N-(N-2-pyridyl-methyleneoxy-carbonyl-leucinyl)-amino-3-N-(4-phenoxy-phenyl-sulfonyl-amino-propan-2-one

Following the procedure of Example 213(a), except substituting“2-pyridyl-methyleneoxy carbonyl-leucine” for “Cbz-leucine”, the titlecompound was prepared: MS(ES) M+H⁺=569.

Example 239

Preparation of1-N-(N-4-carboxy-benzoyl-leucinyl)-amino-3-N-(4-phenoxy-phenyl-sulfonyl)-amino-propan-2-one

1-N-(4-carboxymethyl-benzoyl-leucinyl)-amino-3-N-(4-phenoxy-phenyl-sulfonyl)-amino-propan-2-one(0.105 g, 0.176 mmol) was dissolved in MeOH (5 ml) and water (1 ml),then LiOH-hydrate (15 mg, 0.35 mmol) was added and the reaction wasstirred at RT for 1 h. The reaction was diluted with water, acidifiedwith 6N hydrochloric acid (1 ml), then with EtOAc (2×10 ml). Thecombined organics were dried with magnesium sulfate, filtered,concentrated, chromatographed (silica gel, 50:50:1 EtOAc: hexanes: AcOH)to give the desired product as a white solid (35.6 mg, 35%): MS(ES)M+H⁺=582, M+Na⁺=604.

Example 240

Preparation of1-N-(N-Me-N-Cbz-leucinyl)-amino-3-N-(4-phenoxy-phenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 213(a), except “N-Me-N-Cbz-leucine”for “Cbz-leucine”, the title compound was prepared: MS(ES) M+H⁺=582,M+Na⁺=604, 2M+Na⁺=1185.

Example 241

Preparation of 1-N-(4-phenoxy benzoyl)-amino-3-N-(4-phenoxy-phenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 213(a), except “4-phenoxy benzoicacid” for “Cbz-leucine”, the title compound was prepared: MS(ES)M+H⁺=517, M+Na⁺=539, 2M+Na⁺=1055.

Example 242

Preparation of 1-N-(3-phenoxy-benzoyl)-amino-3-N-(4-phenoxy-phenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 213(a), except “3-phenoxy benzoicacid” for “Cbz-leucine”, the tide compound was prepared: MS(ES) M+H=517,M+Na⁺=539, 2M+Na⁺=1055.

Example 243

Preparation of 1-N-(4phenoxy benzoyl)-amino-3-N-(4-phenoxy-phenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 213(a), except “4-phenoxy benzoicacid” for “Cbz-leucine”, the title compound was prepared: MS(ES)M+H⁺=517, M+Na⁺=539, 2M+Na⁺=1055, M−H⁺=515.

Example 244

Preparation of 1-N-(4biphenylacetyl)-amino-3-N-(4-phenoxy-phenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 213(a), except “4-biphenyl aceticacid” for “Cbz-leucine”, the title compound was prepared: MS(ES)M+H⁺=515, M+Na⁺=537, 2M+Na⁺=1051.

Example 245

Preparation of 1-N-(2-benzyloxybenzoyl)-amino-3-N-(4-phenoxy-phenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 213(a), except “2-benzyloxy- benzoicacid” for “Cbz-leucine”, the title compound was prepared: MS(ES)M+H⁺=531, M+Na⁺=553, 2M+Na⁺=1083.

Example 246

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-(4-benzyloxy-benzoyl)-amino-propan-2-one

Following the procedure of Example 229, except substituting“4-benzyloxy-benzoic acid” for “8-quinoline carboxylic acid “, the titlecompound was prepared: MS(ES) M+H⁺=546, M+Na⁺=568, 2M+Na⁺=1113.

Example 247

Preparation of1-N-(2-(4-biphenyl)₄-methyl-pentamido)-3-N-(4phenoxy-phenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 213(a), except2-(4-biphenyl)-4-methyl-pentanoic acid” for “Cbz-leucine”, the titlecompound was prepared: MS(ES) M+H⁺=571, M+Na⁺=593.

Example 248

Preparation of1-N-(2-(3-biphenyl)₄-methyl-pentamido)-3-N-(4-phenoxy-phenyl-sulfonyl)-amino-propan-2one

a) 3-bromo-phenyl Methyl Acetate

3-Bromo phenyl acetic acid (2.15 g, 10 mmol) was dissolved in ether,then was treated with a solution of diazomethane until the yellow colorpersisted. The reaction was then quenched with AcOH, concentrated invacuo and was used in the next reaction without further purification.

b) 3-biphenyl Methyl Acetate

3-bromo-phenyl methyl acetate (2.29 g, 10 mmol) was dissolved in toluene(30 ml). Then, phenyl boronic acid (1.46 g, 12 mmol) was added followedby aqueous sodium carbonate (2M, 4.24 ml, 40 mmol), thentetrakis(triphenylphosphine) palladium (0.35 g, 0.3 mmol) and wasrefluxed overnight. The reaction was cooled to RT, diluted withsaturated ammonium chloride, then extracted with EtOAc (2×10 ml). Thecombined organics were dried with magnesium sulfate, filtered,concentrated, and chromatographed (silica gel, 5% EtOAc: hexanes) toprovide the desired product as a white solid (1.93 g, 84%): MS(ES):M+H⁺=263.

c) 3-biphenyl Acetic Acid

3-Biphenyl acetyl methyl ester was dissolved in MeOH (40 ml) and water(6 ml), then LiOH-hydrate (0.7 g, 16.8 mmol) was added, and the reactionwas stirred at RT for 2 h. The reaction was diluted with water,acidified with 6N hydrochloric acid (1 ml), then with EtOAc (2×10 ml).The combined organics were dried with magnesium sulfate, filtered, andconcentrated to give the desired product as a white solid (1.66 g, 93%):¹H NMR: d: 7.6-7.25 (m, 9H), 3.7 (s, 2H).

d) 3-(4-biphenyl)-4-methyl-pent-4-enoic Acid

nBuLi (3.26 ml, 1.6 M in hexanes) was added dropwise to a solution ofdiisopropyl amine (0.74 ml, 5.3 mmol) in THF (6 ml) at 0 C. The reactionwas stirred for 15 minutes, then was cooled to −78 C. 3-Biphenyl aceticacid (0.5 g, 2.35 mmol) wasa dissolved in THF (2 ml) and was addeddropwise to the LDA solution. The reaction was warmed to 0 C, stirred 40minutes, then cooled to −78 C. Isobutenyl bromide (0.475 g, 3.52 mmol)was added and the reaction was stirred for 1 h. Water (2 ml) was addedand the THF was removed in vacuo. The reaction was diluted with water,acidified with 6N hydrochloric acid (1 ml), then with EtOAc (2×10 ml).The combined organics were dried with magnesium sulfate, filtered,concentrated, chromatographed (silica gel, 5% MeOH: methylene chloride)to give the desired product as a white solid (1.66 g, 93%): ¹H NMR: d:7.6-7.3 (m, 9H), 4.75 (d, 2H), 3.87 (t, 1H), 2.87 (dd, 1H), 2.50 (dd,1H), 1.70 (s, 3H).

e) 3-(4-biphenyl)-4-methyl-pentanoic Acid

3-(4-Biphenyl)₄-methyl-pent-4-enoic acid (0.5 g, 1.87 mmol) wasdissolved in EtOAc (25 ml). Then, 10% Pd/C (60 mg) was added and thereaction was stirred for 2.5 h under a balloon of hydrogen gas. Thereaction was filtered, concentrated in vacuo, then was redissolved in1:5 EtOAc: EtOH (15 ml). Then, 10% Pd/C (80 mg) was added and thereaction was stirred under a balloon of hydrogen gas overnight. Thereaction was filtered, concentrated in vacuo, and chromatographed(silica gel, 5% MeOH: methylene chloride) to give the desired product asa white solid (1.66 g, 93%): ¹H NMR: d: 7.67.3 (m, 9H), 3.7 (t, 1H),2.07-1.95 (m, 1H), 1.8-1.7 (m, 1H), 1.6-1.45 (m, 1H).

f)1-N-(2-(3-biphenyl)-4-methyl-pentamido)-3-N-(4-phenoxy-phenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 213(a), except3-(4biphenyl)-4-methyl-pentanoic acid” for “Cbz-leucine”, the titlecompound was prepared: MS(ES) M+H⁺=571, M+Na⁺=593.

Example 249

Preparation of 1-N-(3-biphenylacetyl)-amino-3-N-(4phenoxy-phenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 213(a), except “3-biphenyl aceticacid” for “Cbz-leucine”, the title compound was prepared: MS(ES)M+H⁺=515, M+Na⁺=537, 2M+Na⁺=1051.

Example 250

Preparation of 1-N-(NA-pyridylAcetyl-leucinyl)-amino-3-N-(2-benzyloxy-phenyl-sulfonyl)-amino-propan-2-one

a) 1-N-(N-Boc-leucinyl)-amino-3-N-(2-benzyloxyphenyl-sulfonyl)-amino-propan-2-ol

1,3-Diamino-propan-2-ol (3.375 g, 37.5 mmol) was dissolved in DMF (60ml). Then HOBT-hydrate was added (5.5 g, 40.7 mmol), followed byBoc-L-leucine (9.34 g, 37.5 mmol) and EDCI (7.77 g, 40.7 mmol). Thereaction was stirred for 4 h, then diluted with DMF to make a stocksolution of a total volume of 100 ml (0.375 mmol/ml). The stock solution(18 ml, 6.75 mmol) was treated with NMM (0.89 ml, 7.28 mmol), then2-benzyloxy phenyl sulfonyl chloride (1.9 g, 6.72 mmol). The reactionwas stirred an additional 2 h, then was diluted with water, extractedwith EtOAc, dried with magnesium sulfate, filtered, concentrated, andchromatographed (silica gel, 20% EtOAc: hexanes): MS(ES) M+H⁺=550.

b)1-N-(leucinyl)-amino-3-N-(2-benzyloxy-phenyl-sulfonyl)-amino-propan-2-ol

1-N-(Boc-leucinyl)-amino-3-N-(2-benzyloxy phenylsulfonyl)-amino-propan-2-ol (0.7 g, 1.3 mmol) was dissolved in 1:1TFA:DCM (50 ml) and was stirred at RT for 2 h, concentrated in vacuo andwas used in the following reaction without further purification: MS(ES)M+H⁺=450.

c) 1-N-(N-1-pyridylacetyl-leucinyl)-amino-3-N-(2-benzyloxy-phenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 220(b)-(c), except substituting“1-N-leucinyl-amino-3-N-(2-benzyloxy phenyl sulfonyl)-amino-propan-2-ol”for “N-leucinyl-amino-3-N-(4-phenoxy phenylsulfonyl)-amino-propan-2-ol”, the title compound was prepared: MS(ES)M+H⁺=567.

Example 251

Preparation of 1-N-(N-4-pyridylCarbonyl-leucinyl)-amino-3-N-(2-benzyloxy-phenyl-sulfonyl)-amino-propan-2one

Following the procedure of Example 250(a)-(c), except substituting“4-pyridyl carboxylic acid” for “4-pyridyl acetic acid”, the titlecompound was prepared: MS(ES) M+H⁺=553.

Example 252

Preparation of 1-N-(N-4-imidazoleacetic-leucinyl)-amino-3-N-(2-benzyloxy-phenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 250(a)-(c), except substituting“4-imidazole acid” for “4pyridyl-acetic acid”, the title compound wasprepared: MS(ES) M+H⁺=556.

Example 253

Preparation of 1-N-(N-(N,N-dimethylGlycyl)-leucinyl)-amino-3-N-(2-benzyloxy-phenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 250(a)-(c), except substituting“N,N-dimethyl glycine” for “4pyridyl-acetic acid”, the title compoundwas prepared: MS(ES) M+H⁺=533.

Example 254

Preparation of 1-N-(N-(N-methylProlyl-leucinyl)-amino-3-N-(2-benzyloxy-phenyl-sulfonyl)-amino-propan-2-one

Following the procedure of Example 250(a)-(c), except substituting“N-methyl proline” for “4-pyridyl acetic acid”, the title compound wasprepared: MS(ES) M+H⁺=559.

Example 255

Preparation of 1-N-(N-(N-methylpiperidine-4-carbonyl)-leucinyl)-amino-3-N-(2-benzyloxy-phenyl-sulfonyl-amino-propan-2-one

Following the procedure of Example 250(a)-(c), except substituting“N-methyl-piperidine-4-carboxylic acid” for “4-pyridyl acetic acid”, thetitle compound was prepared: MS(ES) M+H⁺=573.

Example 256

Preparation of 1-N-(N-(N-methylpiperidine-4-carbonyl)-leucinyl)-amino-3-N-(2-dibenzofuran-sulfonyl)-amino-propan-2-one

Following the procedure of Example 250(a)-(c), except substituting“N-methyl-piperidine-4-carboxylic acid” for “4pyridyl acetic acid” and“2-dibenzofuran sulfonyl chloride” for “2-benzyloxy phenyl sulphonylchloride” the title compound was prepared: MS(ES) M+H⁺=557.

Example 257

Preparation of 1-N-(N-(N-methylProlyl)-leucinyl)-amino-3-N-(2-dibenzofuran-sulfonyl)-amino-propan-2-one

Following the procedure of Example 250(a)-(c), except substituting“N-methyl proline” for “4-pyridyl acetic acid” and “2-dibenzofuransulfonyl chloride” for “2-benzyloxy phenyl sulphonyl chloride” the tidecompound was prepared: MS(ES) M+H⁺=543.

Example 258

Preparation of 1-N-(N-(N,N-dimethylglycyl)-leucinyl)-amino-3-N-(-(2-dibenzofuran-sulfonyl)-amino-propan-2-one

Following the procedure of Example 250(a)-(c), except substitutingN,N-dimethyl glycine” for “4pyridyl acetic acid” and“2-dibenzofuran-sulfonyl chloride” for “2-benzyloxy phenyl sulphonylchloride” the title compound was prepared: MS(ES) M+H⁺=517.

Example 259

Preparation of 1-N-(N-4-imidazoleacetic-leucinyl)-amino-3-N-(2-dibenzofuran-sulfonyl)-amino-propan-2-one

Following the procedure of Example 250(a)-(c), except substituting“4-imidazole acetic acid” for “4-pyridyl acetic acid” and“2-dibenzofuran sulfonyl chloride” for “2-benzyloxy phenyl sulphonylchloride” the title compound was prepared: MS(ES) M+H⁺=526.

Example 260

Preparation of 1-N-(N-4-pyridylcarbonyl-leucinyl)-amino-3-N-(2-dibenzofuran-sulfonyl)-amino-propan-2-one

Following the procedure of Example 250(a)-(c), except substituting“4-pyridyl carboxlic acid” for “4-pyridyl acetic acid” and“2-dibenzofuran sulfonyl chloride” for “2-benzyloxy phenyl sulphonylchloride” the title compound was prepared: MS(ES) M+H⁺=537.

Example 261

Preparation of 1-N-(N-4-pyridylacetyl-leucinyl)-amino-3-N-2-dibenzofuran-sulfonyl)-amino-propan-2-one

Following the procedure of Example 250(a)-(c), except substituting“2-dibenzofuran sulfonyl chloride” for “2-benzyloxy phenyl sulphonylchloride” the title compound was prepared: MS(ES) M+H⁺=551.

Example 262

Preparation of1-N-(N-4-imidazole-acrylyl-leucinyl)-amino-3-N-(2-dibenzofuran-sulfonyl)-amino-propan-2-one

Following the procedure of Example 250(a)-(c), except substituting“4-imidazole-acryic acid” for “4-pyridyl acetic acid” and“2-dibenzofuran sulfonyl chloride” for “2-benzyloxy phenyl sulphonylchloride” the title compound was prepared: MS(ES) M+H⁺=552.

Example 263

Preparation of1-N-(N-pyrazole-carbonyl-leucinyl)-amino-3-N-(2-dibenzofuran-sulfonyl)-amino-propan-2-one

Following the procedure of Example 250(a)-(c), except substituting“pyrazole carboxylic acid” for “4-pyridyl acetic acid” and“2-dibenzofuran sulfonyl chloride” for “2-benzyloxy phenyl sulphonylchloride” the title compound was prepared: MS(ES) M+H⁺=538.

Example 264

Preparation of1-N-(N-benzoyl-leucinyl)-amino-3-N-(8-quinoline-sulfonyl)-amino-propan-2-one

Following the procedure of Example 250(a)-(c), except substituting“benzoic acid” for “4-pyridyl acetic acid” and “8-quinoline sulfonicacid” for “2-benzyloxy phenyl sulphonyl chloride” the title compound wasprepared: MS(ES) M+H⁺=497.

Example 265

Preparation of1-N-(N-2,5-difluoro-benzoyl-leucinyl)-amino-3-N-(8-quinoline-sulfonyl)-amino-propan-2-one

Following the procedure of Example 250(a)-(c), except substituting“2,5-difluoro-benzoic acid” for “4-pyridyl acetic acid” and “8-quinolinesulfonic acid” for “2-benzyloxy phenyl sulphonyl chloride” the titlecompound was prepared: MS(ES) M+H⁺=535.

Example 266

Preparation of 1-N-(N-2,5-difluoro-phenylacetyl-leucinyl)-amino-3-N-(8-quinoline-sulfonyl)-amino-propan-2-one

Following the procedure of Example 250(a)-(c), except substituting“2,5-difluoro- phenyl acetic acid” for “4-pyridyl acetic acid” and“8-quinoline sulfonic acid” for “2-benzyloxy phenyl sulphonyl chloride”the title compound was prepared: MS(ES) M+H⁺=547.

Example 267

Preparation of 1-N-(N-phenylacetyl-leucinyl)-amino-3-N-(8-quinoline-sulfonyl)-amino-propan-2-one

Following the procedure of Example 250(a)-(c), except substituting“phenyl acetic acid” for “4-pyridyl acetic acid” and “8-quinolinesulfonic acid” for “2-benzyloxy phenyl sulphonyl chloride” the titlecompound was prepared: MS(ES) M+H⁺=511.

Example 268

Preparation of 1-N-(NA-pyridylacetyl-leucinyl)-amino-3-N-(8-quinoline-sulfonyl)-amino-propan-2-one

Following the procedure of Example 250(a)-(c), except substituting“4-pyridyl acetic acid” for “4-pyridyl acetic acid” and “8-quinolinesulfonic acid” for “2-benzyloxy phenyl sulphonyl chloride” the tidecompound was prepared: MS(ES) M+H⁺=512.

Example 269

Preparation of 1-N-(N-pyridylCarbonyl-leucinyl)-amino-3-N-(8-quinoline-sulfonyl)-amino-propan-2-one

Following the procedure of Example 250(a)-(c), except substituting“4-pyridyl carboxylic acid” for “4-pyridyl acetic acid” and “8-quinolinesulfonic acid” for “2-benzyloxy phenyl sulphonyl chloride” the titlecompound was prepared: MS(ES) M+H⁺=498.

Example 270

Preparation of 1-N-(N-4-imidazoleacetyl-leucinyl)-amino-3-N-(8-quinoline-sulfonyl)-amino-propan-2-one

Following the procedure of Example 250(a)-(c), except substituting“4-imidazole acetyl acid” for “4-pyridyl acetic acid” and “8-quinolinesulfonic acid” for “2-benzyloxy phenyl sulphonyl chloride” the titlecompound was prepared: MS(ES) M+H⁺=501.

Example 271

Preparation of 1-N-(N-3-phenylPropionyl-leucinyl)-amino-3-N-(8-quinoline-sulfonyl)-amino-propan-2-one

Following the procedure of Example 250(a)-(c), except substituting“hydrocinnamic acid” for “4-pyridyl acetic acid” and “8-quinolinesulfonic acid” for “2-benzyloxy phenyl sulphonyl chloride” the titlecompound was prepared: MS(ES) M+H⁺=525.

Example 272

Preparation of bis-N,N′-(N-4-pyridyl MethyleneoxyCarbonyl-leucinyl)-1,3-diamino-propan-2-one

Following the procedure of Example 37, except substituting“4-pyridyl-methyleneoxy-carbonyl-leucinyl” for “Cbz-leucine”, the titlecompound was prepared: MS(ES) M+H⁺=585.

Example 273

Preparation of bis-N,N′-(N-3-pyridylmethyleneoxy-carbonyl-leucinyl)-1,3-diamino-propan-2-one

Following the procedure of Example 37, except substituting“3-pyridyl-methyleneoxy-carbonyl-leucinyl” for “Cbz-leucine”, the titlecompound was prepared: MS(ES) M+H⁺=585.

Example 274

Preparation of bis-N,N′-(N-2-pyridyl methyleneoxycarbonyl-leucinyl)-1,3-diamino-propan-2-one

Following the procedure of Example 37, except substituting“2-pyridyl-methyleneoxy-carbonyl-leucinyl” for “Cbz-leucine”, the titlecompound was prepared: MS(ES) M+H⁺=585.

Example 275

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-(2-benzyloxy-benzoyl)-amino propan-2-one

Following the procedure of Example 229, except 2-benzyloxy-benzoic acid”for “8-quinoline-carboxylic acid”, the title compound was prepared:MS(ES) M+H⁺=546.

Example 276

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-(3-benzyloxy-benzoyl)-amino-propan-2-one

Following the procedure of Example 229, except “3-benzyloxy benzoicacid” for “8-quinoline carboxylic acid”, the title compound wasprepared: MS(ES) M+H⁺=546.

Example 277

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-(4-biphenyl-acetyl)-amino-propan-2-one

Following the procedure of Example 229, except “4-biphenyl acetic acid”for “8-quinoline carboxylic acid”, the title compound was prepared:MS(ES) M+H⁺=530.

Example 278

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-(2-carboxymethyl-thiophene-3-sulfonyl)-amino-propan-2-one

Following the procedure of Example 51, except substituting“2-carboxymethyl thiophene-3-sulfonyl” for“4-(3-Chloro-2-cyano-phenoxy)-phenyl sulfonyl chloride”, the titlecompound was prepared: MS(ES) M−H⁺=540.

Example 279

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-methyl-N-(N-Cbz-leucinyl)-amino-propan-2-one

a) N-(Cbz-leucinyl)-amino-propene

N-Cbz-leucine (3.0 g, 11.3 mmol) was dissolved in DMF (50 ml), then NMM(1.3 g, 12.4 mmol) was added, followed by allyl amine (0.65 g, 0.85mmol), and HBTU (4.3 g, 11.3 mmol) and the reaction was stirredovernight at RT. The reaction was diluted with water, extracted withEtOAc, dried with magnesium sulfate, filtered, concentrated, andchromatographed (silica gel, 40% EtOAc: hexanes): MS(ES) M+H⁺=305.

b) N-(Cbz-leucinyl)-amino-propene Oxide

N-(Cbz-leucinyl)-amino-propene (2.95 g, 9.7 mmol) was dissolved inmethylene chloride (100 ml), then mCPBA (5.0 g, 29.1 mmol) was added andthe reaction was stirred overnight. The reaction was diluted withsaturated aqueous sodium bicarbonate, extracted with EtOAc, dried withmagnesium sulfate, filtered, concentrated, and chromatographed (silicagel, 50% EtOAc: hexanes).

c) 1-N-(Cbz-leucinyl)-amino-3-N-methyl-amino-propan-2-ol

N-(Cbz-leucinyl)-amino-propene oxide (400 mg, 1.25 mmol) was dissolvedin isopropanol (5 ml), then aqueous methyl amine (2 ml) was added andthe reaction was heated to 70 C in a sealed bomb for 2 h. The reactionmixture was concentrated in vacuo and was used in the next reactionwithout further purification.

d)1-N-(N-Cbz-leucinyl)-amino-3-N-methyl-N-(N-Cbz-leucinyl)-amino-propan-2-one

Following the procedure of Example 229(a), except substituting“1-N-(Cbz-leucinyl)-amino-3-N-methyl-amino-propan-2-ol” for “8-quinolinecarboxylic acid”, the title compound was prepared: MS(ES) M+H⁺=597.

Example 280

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-methyl-N-(NA-pyridyl-methyloxy-carbonyl-leucinyl-amino-propan-2-one

Following the procedure of Example 279(a)-(d), except substituting “4pyridyl-methyleneoxy-carbonyl leucine” for “Cbz-leucine” in (d), thetitle compound was prepared: MS(ES) M+H⁺=598.

Example 281

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-methyl-N-(2-dibenzofuran-sulfonyl)-amino-propan-2-one

Following the procedure of Example 279(a)-(d), except substituting“2-dibenzofuran sulfonyl chloride” for “Cbz-leucine and HBTU” in (d),the title compound was prepared: MS(ES) M+H⁺=580, M+Na⁺=602.

Example 282

Preparation of1-N-methyl-1-N-(N-Cbz-leucinyl)-amino-3-N-(2-dibenzofuran-sulfonyl)-amino-propan-2-one

Following the procedure of Example 279(a)-(d), except substituting“2-dibenzofuran sulfonyl chloride” for “Cbz-leucine and HBTU” in (a),the title compound was prepared: MS(ES) M+H⁺=580.

Example 283

Preparation of1-N-methyl-1N-(N-Cbz-leucinyl)-amino-3-N-(2-dibenzofuran-sulfonyl)-amino-propan-2-one

Following the procedure of Example 279(a)-(d), except substituting“2-dibenzofuran sulfonyl chloride” for “N-Cbz-leucine” in step (a) and“4-pyridyl methyleneoxy carbonyl-leucine” for “Cbz-leucine” in step (d),the title compound was prepared: MS(ES) M+H⁺=580.

Example 284

Preparation of 1-N-(2-dibenzofuran sulfonyl)-N-methyl)-amino-3N-(N-4-pyridyl-methyleneoxy Carbonyl-leucinyl)-amino-propan-2-one

Following the procedure of Example 279(a)-(d), except substituting “4pyridyl methyl amine” for “allyl amine” and “2-dibenzofuran sulfonylchloride” for “N-Cbz-leucine” in step (a) and “N-4-pyridyl methyleneoxycarbonyl-leucinyl” for “N-Cbz-leucine and HBTU “in step (d), the titlecompound was prepared: MS(ES) M+H⁺=581.

Example 285

Preparation of1-N-(N-Cbz-leucinyl)-amino-3-N-(4-pyridyl-methylene)-3N-(N-Cbz-leucinyl)-amino-propan-2-one

Following the procedure of Example 279(a)-(d), except substituting“4-pyridyl methyl amine” for “methyl amine”, the title compound wasprepared: MS(ES) M+H⁺=674.

Example 286

Preparation of1-N-(Cbz-leucinyl)-amino-3-N-(4-pyridyl-methylene)-3N-(2-dibenzofuranSulfonyl)-amino-propan-2-one

Following the procedure of Example 284(a)-(d), except substituting“2-dibenzofuran sulfonyl chloride” Cbz-leucine and HBTU” in step (d),the title compound was prepared: MS(ES) M+H⁺=657.

Example 287

Preparation of1-N-(Cbz-leucinyl)-amino-3-N-(4-pyridyl-methylene)-3N-(2-dibenzofuransulfonyl)-amino-propan-2-one

Following the procedure of Example 284(a)-(d), except substituting“2-dibenzofuran sulfonyl chloride” Cbz-leucine and HBTU” in step (d),the title compound was prepared: MS(ES) M+H⁺=657.

Example 288

Preparation of 1-N-(4-biphenylacetyl)-amino-3-N-(4-pyridyl-methylene)-3N-(N-Cbz-leucinyl)-amino-propan-2-one

Following the procedure of Example 279(a)-(d), except substituting“4-biphenyl acetic acid” for “Cbz-leucine” in step (a), “4-pyridylmethyl amine” for “methyl amine”, the title compound was prepared:MS(ES) M+H⁺=621.

Example 289

Preparation of1-N-(4-phenoxy-benzoyl)-amino-3-N-(4-pyridyl-methylene)-3N-(N-Cbz-leucinyl)-amino-propan-2-one

Following the procedure of Example 279(a)-(d), except substituting “4phenoxy benzoic acid” for “Cbz-leucine “in step (a), “4-pyridyl methylamine” for “methyl amine”, the title compound was prepared: MS(ES)M+H⁺=623.

Example 290

Preparation of1-N-(2-dibenzofuran-sulfonyl)-amino-3-N-(4-pyridyl-methylene)-3N-(N-Cbz-leucinyl)-amino-propan-2-one

Following the procedure of Example 279(a)-(d), except substituting“2-dibenzofuran sulfonyl chloride” for “Cbz-leucine and HBTU” in step(a),” 4-pyridyl methyl amine” for “methyl amine”, the title compound wasprepared: MS(ES) M+H⁺=657.

Example 291

Preparation of1-N-(N-3-pyridyl-methyleneoxy-carbonyl-leucinyl)-amino-3-N-methyl-N-(2-dibenzofuran-sulfonyl)-amino-propan-2-one

Following the procedure of Example 279(a)-(d), except substituting“N-methyl-N-allyl amine” for “allyl amine” in (a), and“3-pyridyl-methyleneoxy-carbonyl-leucine” for “Cbz-leucine” in step (d),the title compound was prepared: MS(ES) M+H⁺=581.

The above specification and Examples fully disclose how to make and usethe compounds of the present invention. However, the present inventionis not limited to the particular embodiments described hereinabove, butincludes all modifications thereof within the scope of the followingclaims. The various references to journals, patents and otherpublications which are cited herein comprise the state of the art andare incorporated herein by reference as though fully set forth.

We claim:
 1. A compound according to Formula VIII:

wherein: m is 0, 1, or 2; R³² is OCH₂Ar, OCH2C₁₋₆alkyl, aryl substitutedC₀₋₆alkyl, heteroaryl substituted C₀₋₆alkyl, 4-imidazole methylene; 2-,3-, or 4- pyridylmethyleneoxy; 4-pyridyl methylene, 2-pyridyl sulfonyl,4-pyridyl, aryl C₀₋₆alkyloxy, or heteroaryl substituted C₀₋₆alkyloxy;R³³ is C₁₋₆alkyl, —CH₂Ph, or —CH₂CH₂CO₂R³⁴; R³⁴ is —H or C₁₋₆alkyl; R³⁸is Cbz; C₁₋₆alkyl or aryl substituted Cbz; C₁₋₆alkyl —CO; benzoyl;C₁₋₆alkyl or aryl substituted benzoyl; R³⁹ is

Cbz-leucinyl-; 2-, 3-, or 4-pyridyl methyloxycarbonyl-leucinyl-;4-imidazole acetyl-leucinyl-, phenyl acetyl-leucinyl,N,N-dimethyl-glycinyl leucinyl, 4-pyridyl acetyl-leucinyl, 2-pyridylsulfonyl-leucinyl, 4-pyridyl carbonyl-leucinyl, acetyl-leucinyl,benzoyl-leucinyl, 4-phenoxy-benzoyl-, 2- or 3-benzyloxybenzoyl-,biphenyl acetyl, alpha- isobutyl-biphenyl acetyl, Cbz-phenylalaninyl,Cbz-norleucinyl-, Cbz-norvalinyl-, Cbz-glutamyl-, Cbz-epsilon-(t-butylester)-glutamyl; acetyl-leucinyl-, 6- or 8- quinoline carbonyl, biphenylacetyl, alpha- isobutyl-biphenyl acetyl, acetyl, benzoyl, 2- or3-benzyloxy benzoyl, 4-phenoxy benzoyl-, Cbz-amino acid-; 2-,3-, or4-pyridylmethyloxycarbonyl-aminoacid-; aryl C₀-C₆alkyloxy carbonyl-aminoacid-, heteroaryl C₀-C₆alkyloxy carbonyl-amino acid-, aryl C₀-C₆alkyloxycarbonyl-amino acid-, heteroaryl C₀-C₆alkyloxy carbonyl-amino acid-,C₁-C₆alkyloxy carbonyl-amino acid-; C₁-C₆alkyl carbonyl, aryl C₀-C₆alkylcarbonyl, heteroaryl C₀-C₆alkyl carbonyl, aryl C₀-C₆alkyl carbonyl,heteroaryl C₀-C₆alkyl carbonyl, C₁-C₆alkyl sulfonyl, aryl C₀-C₆alkylsulfonyl, heteroaryl C₀-C₆alkyl sulfonyl, aryl C₀-C₆alkyl sulfonyl, orheteroaryl C₀-C₆alkyl sulfonyl; R⁴⁰ is H or C₁₋₆alkyl; R⁴¹ is H orC₁₋₆alkyl; R⁴² is C₁₋₆alkyl, aryl substituted C₁₋₆alkyl, or heteroarylsubstituted C₁₋₆alkyl; provided that R⁴² is H when R⁴³ is C₁₋₆alkyl,aryl substituted C₁₋₆alkyl, or heteroaryl substituted C₁₋₆alkyl; R⁴³ isC₁₋₆alkyl; aryl substituted C₁₋₆alkyl, or hetero aryl substitutedC₁₋₆alkyl; provided that R⁴³ is H when R⁴² is C₁₋₆alkyl, arylsubstituted C₁₋₆alkyl; or heteroaryl substituted C₁₋₆alkyl; andpharmaceutically acceptable salts, hydrates, or solvates thereof.
 2. Acompound according to claim 1 wherein R³⁹ is 2-dibenzofuranylsulfonyl.3. A pharmaceutical composition comprising a compound according to claim1 and a pharmaceutically acceptable carrier, diluent or excipient.
 4. Amethod of inhibiting a cysteine protease comprising administering to apatient in need thereof an effective amount of a compound according toclaim
 1. 5. A method according to claim 4 wherein said cysteine proteaseis cathepsin K.
 6. A method of treating a disease characterized by boneloss comprising inhibiting said bone loss by administering to a patientin need thereof an effective amount of a compound according to claim 1.7. A method according to claim 6 wherein said disease is osteoporosis.8. A method according to claim 6 wherein said disease is periodontitis.9. A method according to claim 6 wherein said disease is gingivitis. 10.A method of treating a disease characterized by excessive cartilage ormatrix degradation comprising inhibiting said excessive cartilage ormatrix degradation by administering to a patient in need thereof aneffective amount of a compound according to claim
 1. 11. A methodaccording to claim 10 wherein said disease is osteoarthritis.
 12. Amethod according to claim 10 wherein said disease is rheumatoidarthritis.